Agonists of guanylate cyclase and their uses

ABSTRACT

This invention provides novel guanylate cyclase C (GC-C) agonists and their therapeutic use. The agonists may be used either alone or in combination with one or more additional agents.

RELATED APPLICATIONS

This application is a continuation of U.S. Utility application Ser. No.14/207,753, filed Mar. 13, 2014, and claims priority to, and benefit of,the U.S. Provisional Application No. 61/790,266, filed on Mar. 15, 2013and the U.S. Provisional Application No. 61/826,749, filed on May 23,2013, the contents of each of which are incorporated herein by referencein their entireties.

INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING

The contents of the text file named “SYPA-014_C01US_SeqList_ST25”, whichwas recorded on Jun. 14, 2017 and is 55 KB in size, are herebyincorporated by reference in their entireties.

FIELD OF THE INVENTION

The present invention relates to novel guanylate cyclase C (GC-C)agonists and their therapeutic use. The agonists may be used eitheralone or in combination with one or more additional agents.

BACKGROUND OF THE INVENTION

Irritable bowel syndrome (IBS) is a common chronic disorder of theintestine that affects 20 to 60 million individuals in the US alone(Lehman Brothers, Global Healthcare-Irritable Bowel Syndrome IndustryUpdate, September 1999). IBS is the most common disorder diagnosed bygastroenterologists (28% of patients examined) and accounts for 12% ofvisits to primary care physicians (Camilleri 2001 Gastroenterology120:652-668).

Diabetes mellitus (DM) is a group of metabolic diseases characterized byhyperglycemia, resulting from defects in insulin secretion, insulinaction, or both. The chronic hyperglycemia of diabetes is associatedwith long-term damage, dysfunction, and failure of various organs,especially the eyes, kidneys, nerves, heart, and blood vessels.Well-known risk factors of type 2 DM are family history, obesity, age,race, prediabetes [impaired fasting glucose (IFG) and/or impairedglucose tolerance (IGT)], gestational DM, and polycystic ovariansyndrome. An association between insulin resistance and inflammation hasbeen reported. Data also indicated a correlation that prediabetes wascommon in patients with IBS, which suggested that the chronicinflammation process might be responsible for the progression to DM.

Hypercholesterolemia has been recognized as a major risk factor forcoronary heart disease (CHD). In clinical trials, reducing serum LDLcholesterol has been demonstrated to decrease the incidence of CHD andto reverse atherosclerotic lesions. Two main classes of clinicallyuseful hypocholesterolemic agents are the 3-hydroxy-3-methylglutarylcoenzyme A (HMG-CoA) reductase inhibitors (e.g., statins) and the bileacid sequestrants. Both induce hepatic LDL receptor activity byincreasing hepatic cholesterol demand. Because the major determinant ofserum cholesterol level is hepatic LDL receptor activity (38), theseagents may share a common mechanism leading to reduction in serumcholesterol.

The guanylate cyclase-C (GC-C) receptor (reviewed by Lucas et al. 2000Pharmacol. Rev 52:375-414 and Vaandrager et al. 2002 Molecular andCellular Biochemistry 230:73-83) is a key regulator in mammals ofintestinal function (although low levels of GC-C have been detected inother tissues). GC-C responds to the endogenous hormones, guanylin anduroguanylin, and to enteric bacterial polypeptides from the heat stableenterotoxin family (ST polypeptides). When an agonist binds to GC-C,there is an elevation of the second messenger, cyclic GMP, and anincrease in chloride and bicarbonate secretion, resulting in an increasein intestinal fluid secretion.

Given the prevalence of diseases associated with GI inflammation,hypercholesterolemia, obesity and inflammatory conditions, there existsa need to develop compositions and methods for effective treatment.

SUMMARY OF THE INVENTION

The present invention provides novel peptides.

The present invention also provides a pharmaceutical composition thatincludes a peptide described herein in a therapeutically effectiveamount and a pharmaceutical carrier, excipient or diluent.

In some embodiments, the peptide increases cGMP production in a cell.

In some embodiments, the peptide is a bicyclic peptide.

The present invention also provides a composition that includes an inertcarrier coated with peptides of the invention and an enteric coatingthat releases the peptide at pH5.0 or pH7.0. The inert carrier includes,for example, a selected from mannitol, lactose, a microcrystallinecellulose or starch.

The present invention further provides a method for treating a conditionthat responds to enhanced cGMP levels in a subject by administering tothe subject a therapeutically effective amount of a peptide of theinvention, and the peptide is administered in an amount sufficient toincrease water transport in the gastrointestinal tract and induce cGMPproduction in a gastrointestinal epithelial cell.

The present invention further provides a method for preventing ortreating a condition that includes, for example, ulcerative colitis,Crohn's disease, irritable bowel syndrome (IBS), non-ulcer dyspepsia,chronic intestinal pseudo-obstruction, functional dyspepsia, colonicpseudo-obstruction, duodenogastric reflux, constipation, chronicconstipation, constipation associated with use of opiate pain killers,post-surgical constipation, constipation associated with neuropathicdisorders, gastroesophageal reflux disease (GERD), Celiac disease,gastroparesis, heartburn, poor gastrointestinal motility, congestiveheart failure, hypertension, benign prostatic hyperplasia (BPH), coloncancer, lung cancer, bladder cancer, liver cancer, salivary gland canceror skin cancer, bronchitis, tissue inflammation, organ inflammation,respiratory inflammation, asthma, COPD, lipid metabolism disorders,biliary disorders, cardiovascular disease, obesity or an endocrinedisorder, by administering to a subject in need thereof atherapeutically effective amount of a peptide of the invention.

The present invention further provides a method of colonic cleansing byadministering to a subject in need thereof an effective amount of apeptide of the invention. The method may further include a step ofadministering to the subject an effective amount of L-glucose,lubiprostone (Amitiza), prucalopride, an agent for treating chronicconstipation, or any combination thereof.

The present invention further provides a method of increasing cGMPproduction in a cell by contacting the cell with a peptide of theinvention.

The present invention provides a use of a peptide described herein inthe manufacture of a medicament for the treatment of a human disease.

Any methods and uses described herein may further include a step ofadministering a therapeutically effective amount of a cGMP-dependentphosphodiesterase inhibitor. The cGMP-dependent phosphodiesteraseinhibitor is administered either concurrently or sequentially with thepeptide. The cGMP-dependent phosphodiesterase inhibitor includes, forexample, sulindac sulfone, zaprinast, motapizone, vardenafil, andsildenafil.

Any methods and uses described herein may further include a step ofadministering a therapeutically effective amount of at least oneanti-inflammatory agent. The anti-inflammatory agent is a steroid ornonsteroid anti-inflammatory drug (NSAID).

The peptide of the invention may have the sequence of any one of Table1.

In some embodiments, the peptide isAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶(SEQ ID NO: 1),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶(SEQ ID NO: 32),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Thr¹⁶(SEQ ID NO: 119),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶(SEQ ID NO: 120),dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶(SEQ ID NO: 17), orpyGlu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶(SEQ ID NO: 56).

Other features and advantages of the invention will be apparent from andare encompassed by the following detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the stimulation of cyclic GMP synthesis bySP-304 and SP-304 with α-aminoadipic acid in the 3^(rd) position fromthe N-terminus.

DETAILED DESCRIPTION

The present invention is based upon the development of novel agonists ofguanylate cyclase-C (GC-C). The agonists are plecanatide derivatives,analogs of uroguanylin, guanylin, lymphoguanylin and ST peptide and havesuperior properties such as higher activity or potency. Particularly,these analogs contain an α-aminoadipic acid (Aad), preferably at the3^(rd) position from the N-terminus of each peptide or at the positionto the N-terminal side next to the first cysteine (“Cys”) residue.

The gualylate cyclase-C agonists according to the invention includeamino acid sequences represented by Formulae I-Aad, II-Aad, III-Aad,IV-Aad, V-Aad, VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aador XXI-Aad as well as those amino acid sequences summarized below inTable 1. The gualylate cyclase-C agonists according to the invention arecollectively referred to herein as “Aad-GCRA peptides”.

TABLE 1 Alpha-aminoadipic acid derivatives of GCRA Peptides Position ofCorresponds to: Disulfide bond Structure SEQ ID NO SP-304 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶  1 SP-326 C3:C11, C6:C14Asp¹-Aad²-Cys³-Glu⁴-Leu⁵-Cys⁶-Val⁷-Asn⁸-Val⁹-Ala¹⁰-Cys¹¹-Thr¹²-Gly¹³-Cys¹⁴-Leu^(l5)  2 SP-327 C3:C11, C6:C14Asp¹-Aad²-Cys³-Glu⁴-Leu⁵-Cys⁶-Val⁷-Asn⁸-Val⁹-Ala¹⁰-Cys¹¹-Thr¹²-Gly¹³-Cys¹⁴  3 SP-328 C2:C10, C5:C13Aad¹-Cys²-Glu³-Leu⁴-Cys⁵-Val⁶-Asn⁷-Val⁸-Ala⁹-Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³-Leu^(l4)  4 SP-329 C2:C10, C5:C13Aad¹-Cys²-Glu³-Leu⁴-Cys⁵-Val⁶-Asn⁷-Val⁸-Ala⁹-Cys¹⁰-Thr¹¹-Gly¹²-Cys¹³   5SP-332 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶  6 SP-333 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶  7 SP-334 C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶  8 SP-336 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶  9 SP-337 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys4-Glu⁵-dLeu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 10 SP-338 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys4-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵ 11 SP-342 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 12 SP-343 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 13 SP-344 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 14 SP-347 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 15 SP-348 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 16 SP-350 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 17 SP-352 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 18 SP-359 C4:C12, C7:C15PEG3-dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 19 SP-360 C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 20 SP-368 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dNal¹⁶ 21 SP-369 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-AIB⁸-Asn⁹-AIB¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 22 SP-370 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Asp[Lactam]⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Orn¹⁵-dLeu¹⁶ 23 SP-371 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 24 SP-372 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 25 N1 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 26 N2 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 27 N3 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Tyr⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶PEG3 28 N4 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 29 N5 C4:C12, C7:C15PEG3-dAsn¹-Asp²-Aad³-Cys⁴-Glu5-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶ 30 N6 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Ser⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶-PEG3 31 N7 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 32 N8 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶-PEG3 33 N9 C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 34 N10 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶-PEG3 35 N11 C4:C12, C7:C15PEG³-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶-PEG3 36 N12 C4:C12, C7:C15PEG³-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶ 37 N13 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶-PEG3 38 Formula I C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ 39 (I-Aad) Formula II C4:C12, C7:C15Xaa_(n1)-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa_(n2)¹⁶  40 (II-Aad) Formula III 4:12, 7:15Xaa_(n1)-Maa⁴-Glu⁵-Xaa⁶-Maa⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Maa¹²-Thr¹³-Gly¹⁴-Maa¹⁵-Xaa_(n2) 41 (III-Aad) Formula IV 4:12, 7:15Xaa_(n1)-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵-Xaa_(n2) 42 (IV-Aad) Formula V C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-Xaa¹⁶ 43 (V-Aad) Formula VI C4:C12, C7:C15dAsn¹-Glu²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ 44 (VI-Aad) Formula VII-a C4:C12, C7:C15dAsn¹-dGlu²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ 45 (VI-a-Aad) Formula VII-b C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Asn⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ 46 (VI-b-Aad) Formula VIII C4:C12, C7:C15dAsn¹-dAsp²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Tyr⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ 47 (VIII-Aad) Formula C4:C12, C7:C15dAsn¹-dGlu²-Aad³-Cys⁴-Xaa⁵-Xaa⁶-Cys⁷-Xaa⁸-Tyr⁹-Xaa¹⁰-Xaa¹¹-Cys¹²-Xaa¹³-Xaa¹⁴-Cys¹⁵-d-Xaa¹⁶ 48 IX (IX-Aad) Formula XXI C4:C12, C7:C15Xaa_(n1)-Cys⁴-Xaa⁵-Xaa⁶-Xaa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹⁻Cys¹²-Xaa¹³-Xaa¹⁴-Xaa¹⁵-Xaa_(n2)¹⁶  49 (XXI-Aad) SP-363 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu-AMIDE¹⁶ 50 SP-364 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer¹⁶ 51 SP-365 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dSer-AMIDE¹⁶ 52 SP-366 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr¹⁶ 53 SP-367 C4:C12, C7:C15dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dTyr-AMIDE¹⁶ 54 SP-373 C4:C12, C7:C15Pyglu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu-AMIDE¹⁶ 55 / C4:C12, C7:C15Pyglu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 56 SP-304diPEG C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶-PEG3 57 SP-304N-PEG C4:C12, C7:C15PEG3-Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 58 SP-304C-PEG C4:C12, C7:C15Asn¹-Asp²-Glu³-Cys⁴-Aad⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶PEG3 59 Formula XVIII C4:C12, C7:C15Xaa¹-Xaa²-Aad³-Maa⁴-Xaa⁵-Xaa⁶-Maa⁷-Xaa⁸-Xaa⁹-Xaa¹⁰-Xaa¹¹-Maa¹²-Xaa¹³-Xaa¹⁴-Maa¹⁵-Xaa¹⁶ 60 (XVIII-Aad) N32 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 61 N34 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 62 N36 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 63 N38 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 64 N40 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 65 N42 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 66 N44 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 67 N46 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 68 N48 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 69 N50 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 70 N52 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 71 N54 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 72 N56 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 73 N58 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 74 N60 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 75 N62 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 76 N65 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 77 N67 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 78 N69 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 79 N71 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 80 N73 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 81 N75 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 82 N77 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 83 N79 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶ 84 N81 C4:C12, C7:C15Glu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 85 N83 C4:C12, C7:C15Glu¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 86 N85 C4:C12, C7:C15Asp¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 87 N87 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 88 N88 C4:C12, C7:C15Asp¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 89 N89 C4:C12, C7:C15Gln¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 90 N91 C4:C12, C7:C15Gln¹-Glu²-Asp³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 91 N92 C4:C12, C7:C15Gln¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 92 N93 C4:C12, C7:C15Lys¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 93 N95 C4:C12, C7:C15Lys¹-Glu²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Ile⁸-Asn⁹-Met¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ 94 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Thr¹⁶119 C4:C12, C7:C15Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶120

The Aad-GCRA peptides described herein bind the guanylate cyclase C(GC-C) and stimulate intracellular production of cyclic guanosinemonophosphate (cGMP). Optionally, the Aad-GCRA peptides induceapoptosis. In some aspects, the Aad-GCRA peptides stimulateintracellular cGMP production at higher levels than naturally occurringGC-C agonists (e.g., uroguanylin, guanylin, lymphoguanylin and STpeptides) and/or SP-304.

For example, the Aad-GCRA peptides of the invention stimulate 5, 10%,20%, 30%, 40%, 50%, 75%, 90% or more intracellular cGMP compared tonaturally occurring GC-C agonists and/or SP-304. The terms induced andstimulated are used interchangeably throughout the specification.

The Aad-GCRA peptides described herein have therapeutic value in thetreatment of a wide variety of disorders and conditions including forexample lipid metabolism disorders, biliary disorders, gastrointestinaldisorders, inflammatory disorders, lung disorders, cancer, cardiacdisorders including cardiovascular disorders, eye disorders, oraldisorders, blood disorders, liver disorders, skin disorders, prostatedisorders, endocrine disorders, increasing gastrointestinal motility andobesity. Lipid metabolism disorders include, but not limited to,dyslipidemia, hyperlipidemia, hypercholesterolemia,hypertriglyceridemia, sitosterolemia, familial hypercholesterolemia,xanthoma, combined hyperlipidemia, lecithin cholesterol acyltransferasedeficiency, tangier disease, abetalipoproteinemia, erectile dysfunction,fatty liver disease, and hepatitis. Billary disorders includegallbladder disorders such as for example, gallstones, gall bladdercancer cholangitis, or primary sclerosing cholangitis; or bile ductdisorders such as for example, cholecystitis, bile duct cancer orfascioliasis. Gastointestinal disorders include for example, irritablebowel syndrome (IBS), non-ulcer dyspepsia, chronic intestinalpseudo-obstruction, functional dyspepsia, colonic pseudo-obstruction,duodenogastric reflux, gastroesophageal reflux disease (GERD), ileusinflammation (e.g., post-operative ileus), gastroparesis, heartburn(high acidity in the GI tract), constipation (e.g., chronicconstipation; constipation associated with use of medications such asopioids, osteoarthritis drugs, osteoporosis drugs; post surgicalconstipation, constipation associated with neuropathic disorders).Inflammatory disorders include tissue and organ inflammation such askidney inflammation (e.g., nephritis), gastrointestinal systeminflammation (e.g., Crohn's disease and ulcerative colitis); necrotizingenterocolitis (NEC); pancreatic inflammation (e.g., pancreatis), lunginflammation (e.g., bronchitis or asthma) or skin inflammation (e.g.,psoriasis, eczema). Lung Disorders include for example chronicobstructive pulmonary disease (COPD), and fibrosis. Cancer includestissue and organ carcinogenesis including metatases such as for examplegastrointestinal cancer, (e.g., gastric cancer, esophageal cancer,pancreatic cancer colorectal cancer, intestinal cancer, anal cancer,liver cancer, gallbladder cancer, or colon cancer; lung cancer; thyroidcancer; skin cancer (e.g., melanoma); oral cancer; urinary tract cancer(e.g., bladder cancer or kidney cancer); blood cancer (e.g. myeloma orleukemia) or prostate cancer. Cardiac disorders include for example,congestive heart failure, trachea cardia hypertension, high cholesterol,or high tryglycerides. Cardiovascular disorders include for exampleaneurysm, angina, atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, Aad derivates of GC-C agonist described herein may also beuseful to facilitate liver regeneration in liver transplant patients.Eye disorders include for example increased intra-ocular pressure,glaucoma, dry eyes retinal degeneration, disorders of tear glands or eyeinflammation. Skin disorders include for example xerosis. Oral disordersinclude for example dry mouth (xerostomia), Sjögren's syndrome, gumdiseases (e.g., periodontal disease), or salivary gland duct blockage ormalfunction. Prostate disorders include for example benign prostatichyperplasia (BPH). Endocrine disorders include for example diabetesmellitus, hyperthyroidism, hypothyroidism, and cystic fibrosis.

As used herein, the term “guanylate cyclase receptor (GCR)” refers tothe class of guanylate cyclase C receptor on any cell type to which theinventive agonist peptides or natural agonists described herein bind. Asused herein, “intestinal guanylate cyclase receptor” is foundexclusively on epithelial cells lining the GI mucosa. Uroguanylin,guanylin, and ST peptides are expected to bind to these receptors andmay induce apoptosis. The possibility that there may be differentreceptors for each agonist peptide is not excluded. Hence, the termrefers to the class of guanylate cyclase receptors on epithelial cells.

As used herein, the term “GCR agonist” is meant to refer to peptidesand/or other compounds that bind to an intestinal guanylate cyclasereceptor and stimulate fluid and electrolyte transport. This term alsocovers fragments and pro-peptides that bind to GCR and stimulate fluidand water secretion.

As used herein, the term “substantially equivalent” is meant to refer toa peptide that has an amino acid sequence equivalent to that of thebinding domain where certain residues may be deleted or replaced withother amino acids without impairing the peptide's ability to bind to anintestinal guanylate cyclase receptor and stimulate fluid andelectrolyte transport.

Addition of carriers (e.g., phosphate-buffered saline or PBS) and othercomponents to the composition of the present invention is well withinthe level of skill in this art. In addition to the compound, suchcompositions may contain pharmaceutically acceptable carriers and otheringredients known to facilitate administration and/or enhance uptake.Other formulations, such as microspheres, nanoparticles, liposomes, andimmunologically-based systems may also be used in accordance with thepresent invention. Other examples include formulations with polymers(e.g., 20% w/v polyethylene glycol) or cellulose, or entericformulations.

Without intending to be bound by any theory, it is envisioned that iontransport across the plasma membrane may prove to be an importantregulator of the balance between cell proliferation and apoptosis thatwill be affected by agents altering cGMP concentrations. Uroguanylin hasbeen shown to stimulate K+ efflux, Ca++ influx and water transport inthe gastrointestinal tract (3). Moreover, atrial natriuretic peptide(ANP), a peptide that also binds to a specific guanylate cyclasereceptor, has also been shown to induce apoptosis in rat mesangialcells, and to induce apoptosis in cardiac myocytes by a cGMP mechanism(21-24).

Binding of the present agonists to a guanylate cyclase receptorstimulates production of cGMP. This ligand-receptor interaction, viaactivation of a cascade of cGMP-dependent protein kinases and CFTR,induces apoptosis in target cells. Therefore, administration of thenovel peptides defined by Formulae I-Aad, II-Aad, III-Aad, IV-Aad,V-Aad, VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad orXXI-Aad and those listed in Table 1 are useful in eliminating or, atleast retarding, the onset of lipid metabolism disorders, biliarydisorders, gastrointestinal disorders, inflammatory disorders, lungdisorders, cancer, cardiac disorders including cardiovascular disorders,eye disorders, oral disorders, blood disorders, liver disorders, skindisorders, prostate disorders, endocrine disorders, increasinggastrointestinal motility and obesity. Lipid metabolism disordersinclude, but not limited to, dyslipidemia, hyperlipidemia,hypercholesterolemia, hypertriglyceridemia, sitosterolemia, familialhypercholesterolemia, xanthoma, combined hyperlipidemia, lecithincholesterol acyltransferase deficiency, tangier disease,abetalipoproteinemia, erectile dysfunction, fatty liver disease, andhepatitis. Billary disorders include gallbladder disorders such as forexample, gallstones, gall bladder cancer cholangitis, or primarysclerosing cholangitis; or bile duct disorders such as for example,cholecystitis, bile duct cancer or fascioliasis. Gastointestinaldisorders include for example, irritable bowel syndrome (IBS), non-ulcerdyspepsia, chronic intestinal pseudo-obstruction, functional dyspepsia,colonic pseudo-obstruction, duodenogastric reflux, gastroesophagealreflux disease (GERD), ileus inflammation (e.g., post-operative ileus),gastroparesis, heartburn (high acidity in the GI tract), constipation(e.g., chronic constipation, constipation associated with use ofmedications such as opioids, osteoarthritis drugs, osteoporosis drugs;post surgical constipation, constipation associated with neuropathicdisorders). Inflammatory disorders include tissue and organ inflammationsuch as kidney inflammation (e.g., nephritis), gastrointestinal systeminflammation (e.g., Crohn's disease and ulcerative colitis); necrotizingenterocolitis (NEC); pancreatic inflammation (e.g., pancreatis), lunginflammation (e.g., bronchitis or asthma) or skin inflammation (e.g.,psoriasis, eczema). Lung Disorders include for example chronicobstructive pulmonary disease (COPD), and fibrosis. Cancer includestissue and organ carcinogenesis including metatases such as for examplegastrointestinal cancer, (e.g., gastric cancer, esophageal cancer,pancreatic cancer colorectal cancer, intestinal cancer, anal cancer,liver cancer, gallbladder cancer, or colon cancer; lung cancer; thyroidcancer; skin cancer (e.g., melanoma); oral cancer; urinary tract cancer(e.g. bladder cancer or kidney cancer); blood cancer (e.g. myeloma orleukemia) or prostate cancer. Cardiac disorders include for example,congestive heart failure, trachea cardia hypertension, high cholesterol,or high tryglycerides. Cardiovascular disorders include for exampleaneurysm, angina, atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, Aad derivates of GC-C agonist described herein may also beuseful to facilitate liver regeneration in liver transplant patients.Eye disorders include for example increased intra-ocular pressure,glaucoma, dry eyes retinal degeneration, disorders of tear glands or eyeinflammation. Skin disorders include for example xerosis. Oral disordersinclude for example dry mouth (xerostomia), Sjögren's syndrome, gumdiseases (e.g., periodontal disease), or salivary gland duct blockage ormalfunction. Prostate disorders include for example benign prostatichyperplasia (BPH). Endocrine disorders include for example diabetesmellitus, hyperthyroidism, hypothyroidism, and cystic fibrosis.

AAD-GCRA Peptides

In one aspect, the invention provides an Aad-GCRA peptide. The Aad-GCRApeptides are analogues uroguanylin, guanylin, lymphoguanylin and STpeptides. No particular length is implied by the term “peptide”.Particularly, these analogs contain an α-aminoadipic acid (Ad),preferably at the 3rd position from the N-terminus of each peptide or atthe position to the N-terminal side next to the first cysteine (“Cys)residue. In some embodiments, the Aad-GCRA peptide is less than 25 aminoacids in length, e.g., less than or equal to 20, 19, 18, 17, 16, 15, 14,13, 12, 11, 10, or 5 amino acid in length.

The Aad-GCRA peptides can be polymers of L-amino acids, D-amino acids,or a combination of both. For example, in various embodiments, thepeptides are D retro-inverso peptides. The term “retro-inverso isomer”refers to an isomer of a linear peptide in which the direction of thesequence is reversed and the chirality of each amino acid residue isinverted. See, e.g., Jameson et al., Nature, 368, 744-746 (1994); Bradyet al., Nature, 368, 692-693 (1994). The net result of combiningD-enantiomers and reverse synthesis is that the positions of carbonyland amino groups in each amide bond are exchanged, while the position ofthe side-chain groups at each alpha carbon is preserved. Unlessspecifically stated otherwise, it is presumed that any given L-aminoacid sequence of the invention may be made into a D retro-inversopeptide by synthesizing a reverse of the sequence for the correspondingnative L-amino acid sequence. For example an Aad-GCRA peptide includesthe sequence defined by Formulae I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad,VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad andthose listed in Table 1.

By inducing cGMP production is meant that the Aad-GCRA peptide inducesthe production of intracellular cGMP. Intracellular cGMP is measured bymethods known in the art. For example, the Aad-GCRA peptide of theinvention stimulate 5%, 10%, 20%, 30%, 40%, 50%, 75%, 90% or moreintracellular cGMP compared to naturally occurring GC-C agonists.Optionally, the Aad-GCRA peptides of the invention of the inventionstimulate 5%, 10%, 20%, 30%, 40%, 50%, 75%, 90% or more intracellularcGMP compared SP-304. In further embodiments, the Aad-GCRA peptidestimulates apoptosis, e.g., programmed cell death or activates thecystic fibrosis transmembrane conductance regulator (CFTR).

As used herein PEG3, 3 PEG, is meant to denote polyethylene glycol suchas include aminoethyloxy-ethyloxy-acetic acid (AeeA).

As used herein, the term “AMIDE” is meant to denote that the terminalcarboxylic acid is replaced with an amide group, i.e., the terminal COOHis replaced with CONH₂.

As used herein, the term “pyGlu” refers to pyroglutamic acid.

As used herein (e.g., in Formulae I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad,VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad),X_(aa) is any natural, unnatural amino acid or amino acid analogue;M_(aa) is a Cysteine (Cys), Penicillamine (Pen) homocysteine, or3-mercaptoproline.

Xaa_(n1) is meant to denote an amino acid sequence of any any natural,unnatural amino acid or amino acid analogue that is one, two or threeresidues in length. In some embodiments, when Xaa_(n1) represents oneamino acid, Xaa_(n1) is an α-aminoadipic acid (Aad). In someembodiments, when Xaa_(n1) represents two amino acids, the secondresidue from the N-terminus is an α-aminoadipic acid (Aad). In someembodiments, when Xaa_(n1) represents three amino acids, the thirdresidue from the N-terminus is an α-aminoadipic acid (Aad).

Xaa_(n2) is meant to denote an amino acid sequence of any any natural,unnatural amino acid or amino acid analogue that is zero or one residuein length; and Xaa_(n3) is meant to denote an amino acid sequence of anyany natural, unnatural amino acid or amino acid analogue that is zero,one, two, three, four, five or six residues in length. Additionally, anyamino acid represented by Xaa, may be an L-amino acid, a D-amino acid, amethylated amino acid, a florinated amino acid or any combination ofthereof. Preferably the amino acids at the N-terminus, C-terminus orboth are D-amino acids. Optionally, any Aad-GCRA peptide represented byFormulae I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad, VII-a-Aad,VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad may contain on or morepolyethylene glycol residues at the N-terminus, C-terminus or both. Anexemplary polyethylene glycol includes aminoethyloxy-ethyloxy-aceticacid and polymers thereof. In some embodiments, any Aad-GCRA peptiderepresented by Formulae I-Aad, II-Aad, III-Aad, IV-Aad, V-Aad, VI-Aad,VII-a-Aad, VII-b-Aad, VIII-Aad, IX-Aad, XVIII-Aad or XXI-Aad may containAMIDE at the C-terminus.

Specific examples of Aad-GCRA peptides that can be used in the methodsand formulations of the invention include a peptide selected from Table1.

In some embodiments, Aad-GCRA peptides include peptides having the aminoacid sequence of Formula I-Aad. In some embodiments, at least one aminoacid of Formula I-Aad is a D-amino acid or a methylated amino acidand/or the amino acid at position 16 is a serine. Preferably, the aminoacid at position 16 of Formula I-Aad is a D-amino acid or a methylatedamino acid. For example, the amino acid at position 16 of Formula I-Aadis a d-leucine or a d-serine. Optionally, one or more of the amino acidsat positions 1-2 of Formula I-Aad are D-amino acids or methylated aminoacids or a combination of D-amino acids or methylated amino acids. Forexample, Asn¹ or Asp² (or a combination thereof) of Formula I-Aad is aD-amino acid or a methylated amino acid. Preferably, the amino acid atposition Xaa⁶ of Formula I-Aad is a leucine, serine or tyrosine.

In alternative embodiments, Aad-GCRA peptides include peptides havingthe amino acid sequence of Formula II-Aad. In some embodiments, at leastone amino acid of Formula II-Aad is a D-amino acid or a methylated aminoacid. Preferably, the amino acid denoted by Xaa_(n2) of Formula II-Aadis a D-amino acid or a methylated amino acid. In some embodiments, theamino acid denoted by Xaa_(n2) of Formula II-Aad is a leucine, ad-leucine, a serine, or a d-serine. Preferably, the one or more aminoacids denoted by Xaa_(n1) of Formula II-Aad are D-amino acids ormethylated amino acids. Preferably, the amino acid at position Xaa⁶ ofFormula II-Aad is a leucine, a serine, or a tyrosine. In someembodiments, when Xaa_(n1) represents one amino acid, Xaa_(n1) is anα-aminoadipic acid (Aad). In some embodiments, when Xaa_(n1) representstwo amino acids, the second residue from the N-terminus is anα-aminoadipic acid (Aad). In some embodiments, when Xaa_(n1) representsthree amino acids, the third residue from the N-terminus is anα-aminoadipic acid (Aad). In some embodiments, Xaa¹ is a pyroglutamicacid. In some embodiments, Xaa² is glutamic acid or d-glutamic acid. Insome embodiments, Xaa⁸ and Xaa¹⁰ are AIB. In some embodiments, Xaa⁹ istyrosine. In some embodiments, Xaa¹⁶ is dNal.

In some embodiments, Aad-GCRA peptides include peptides having the aminoacid sequence of Formula III-Aad. In some embodiments, at least oneamino acid of Formula III-Aad is a D-amino acid or a methylated aminoacid and/or Maa is not a cysteine. Preferably, the amino acid denoted byXaa_(n2) of Formula III-Aad is a D-amino acid or a methylated aminoacid. In some embodiments the amino acid denoted by Xaa_(n2) of FormulaIII-Aad is a leucine, a d-leucine, a serine, or a d-serine. Preferably,the one or more amino acids denoted by Xaa_(n1) of Formula III-Aad areD-amino acids or methylated amino acids. Preferably, the amino acid atposition Xaa⁶ of Formula III-Aad is a leucine, a serine, or a tyrosine.In some embodiments, when Xaa_(n1) represents one amino acid, Xaa_(n1)is an α-aminoadipic acid (Aad). In some embodiments, when Xaa_(n1)represents two amino acids, the second residue from the N-terminus is anα-aminoadipic acid (Aad). In some embodiments, when Xaa_(n1) representsthree amino acids, the third residue from the N-terminus is anα-aminoadipic acid (Aad). In some embodiments, Xaa¹ is a pyroglutamicacid. In some embodiments, Xaa² is glutamic acid or d-glutamic acid. Insome embodiments, Xaa¹⁶ is dNal.

In other embodiments, Aad-GCRA peptides include peptides having theamino acid sequence of Formula IV-Aad. In some embodiments, at least oneamino acid of Formula IV-Aad is a D-amino acid or a methylated aminoacid, and/or Maa is not a cysteine. Preferably, the Xaa_(n2) of FormulaIV-Aad is a D-amino acid or a methylated amino acid. In someembodiments, the amino acid denoted by Xaa_(n2) of Formula IV-Aad is aleucine, a d-leucine, a serine, or a d-serine. Preferably, the one ormore of the amino acids denoted by Xaa_(n1) of Formula IV-Aad areD-amino acids or methylated amino acids. In some embodiments, whenXaa_(n1) represents one amino acid, Xaa_(n1) is an α-aminoadipic acid(Aad). In some embodiments, when Xaa_(n1) represents two amino acids,the second residue from the N-terminus is an α-aminoadipic acid (Aad).In some embodiments, when Xaa_(n1) represents three amino acids, thethird residue from the N-terminus is an α-aminoadipic acid (Aad).Preferably, the amino acid denoted Xaa⁶ of Formula IV is a leucine, aserine, or a tyrosine. In some embodiments, Xaa¹ is a pyroglutamic acid.In some embodiments, Xaa² is glutamic acid or d-glutamic acid. In someembodiments, Xaa⁸ and Xaa¹⁰ are AIB. In some embodiments, Xaa⁹ istyrosine. In some embodiments, Xaa¹⁶ is dNal.

In further embodiments, Aad-GCRA peptides include peptides having theamino acid sequence of Formula V-Aad. In some embodiments, at at leastone amino acid of Formula V-Aad is a D-amino acid or a methylated aminoacid. Preferably, the amino acid at position 16 of Formula V-Aad is aD-amino acid or a methylated amino acid. For example, the amino acid atposition 16 (i.e., Xaa¹⁶) of Formula V-Aad is a d-leucine or a d-serine.Optionally, one or more of the amino acids at positions 1 and 2 ofFormula V-Aad are D-amino acids or methylated amino acids or acombination of D-amino acids or methylated amino acids. For example,Asn¹ or Asp² (or a combination thereof) of Formula V-Aad is a D-aminoacids or a methylated amino acid. Preferably, the amino acid denoted atXaa⁶ of Formula V-Aad is a leucine, a serine, or a tyrosine.

In additional embodiments, Aad-GCRA peptides include peptides having theamino acid sequence of Formula VI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad orIX-Aad. Preferably, the amino acid at position 6 of Formula VI-Aad,VII-a-Aad, VII-b-Aad, VIII-Aad or IX-Aad is a leucine, a serine, or atyrosine. In some aspects the amino acid at position 16 of FormulaVI-Aad, VII-a-Aad, VII-b-Aad, VIII-Aad or IX-Aad is a leucine or aserine. Preferably, the amino acid at position 16 of Formula VI-Aad,VII-a-Aad, VII-b-Aad, VIII-Aad or IX-Aad is a D-amino acid or amethylated amino acid.

In some embodiments, GCRA peptides include peptides containing the aminoacid sequence of Formula XVIII-Aad. Preferably, the amino acid atposition 1 of Formula XVIII-Aad is a glutamic acid, aspartic acid,glutamine or lysine. Preferably, the amino acid at position 2 and 3 ofFormula XVIII-Aad is a glutamic acid, or an aspartic acid. Preferably,the amino acid at position 5 is a glutamic acid. Preferably, the aminoacid at position 6 of Formula XVIII-Aad is an isoleucine, valine,serine, threonine or tyrosine. Preferably, the amino acid at position 8of Formula XVIII-Aad is a valine or isoleucine. Preferably, the aminoacid at position 9 of Formula XVIII-Aad is an asparagine. Preferably,the amino acid at position 10 of Formula XVIII-Aad is a valine or amethionine. Preferably, the amino acid at position 11 of FormulaXVIII-Aad is an alanine. Preferably, the amino acid at position 13 ofFormula XVIII-Aad is a threonine. Preferably, the amino acid at position14 of Formula XVIII-Aad is a glycine. Preferably, the amino acid atposition 16 of Formula XVIII-Aad is a leucine, serine, threonine ortyrosine.

In some embodiments, GCRA peptides include peptides having the aminoacid sequence of Formula XXI-Aad. In some embodiments, at least oneamino acid of Formula XXI-Aad is a D-amino acid or a methylated aminoacid. Preferably, the amino acid denoted by Xaa_(n2) of Formula XXI-Aadis a D-amino acid or a methylated amino acid. In some embodiments, theamino acid denoted by Xaa_(n2) of Formula XXI-Aad is a leucine, ad-leucine, a serine, or a d-serine. Preferably, the one or more aminoacids denoted by Xaa_(n1) of Formula XXI-Aad are D-amino acids ormethylated amino acids. In some embodiments, when Xaa_(n1) representsone amino acid, Xaa_(n1) is an α-aminoadipic acid (Aad). In someembodiments, when Xaa_(n1) represents two amino acids, the secondresidue from the N-terminus is an α-aminoadipic acid (Aad). In someembodiments, when Xaa_(n1) represents three amino acids, the thirdresidue from the N-terminus is an α-aminoadipic acid (Aad). Preferably,the amino acid at position Xaa⁶ of Formula XXI-Aad is a leucine, aserine, or a tyrosine. In some embodiments, Xaa¹ is a pyroglutamic acid.In some embodiments, Xaa² is glutamic acid or d-glutamic acid. In someembodiments, Xaa⁷ is an aspartic acid and forms a lactam bridge withXaa¹⁵. In some embodiments, Xaa⁸ and Xaa¹⁰ are AIB. In some embodiments,Xaa⁹ is tyrosine. In some embodiments, Xaa¹⁵ is an Orn. In someembodiments, Xaa¹⁶ is dNal.

In certain embodiments, one or more amino acids of the Aad-GCRA peptidescan be replaced by a non-naturally occurring amino acid or a naturallyor non-naturally occurring amino acid analog. There are many amino acidsbeyond the standard 20 (Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His,Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val). Some arenaturally-occurring others are not. (See, for example, Hunt, TheNon-Protein Amino Acids: In Chemistry and Biochemistry of the AminoAcids, Barrett, Chapman and Hall, 1985). For example, an aromatic aminoacid can be replaced by 3,4-dihydroxy-L-phenylalanine,3-iodo-L-tyrosine, triiodothyronine, L-thyroxine, phenylglycine (Phg) ornor-tyrosine (norTyr). Phg and norTyr and other amino acids includingPhe and Tyr can be substituted by, e.g., a halogen, —CH3, —OH, —CH2NH3,—C(O)H, —CH2CH3, —CN, —CH2CH2CH3, —SH, or another group. Any amino acidcan be substituted by the D-form of the amino acid.

With regard to non-naturally occurring amino acids or naturally andnon-naturally occurring amino acid analogs, a number of substitutions inthe polypeptide and agonists described herein are possible alone or incombination.

For example, glutamine residues can be substituted withgamma-Hydroxy-Glu or gamma-Carboxy-Glu. Tyrosine residues can besubstituted with an alpha substituted amino acid such asL-alpha-methylphenylalanine or by analogues such as: 3-Amino-Tyr;Tyr(CH3); Tyr(PO3(CH3)2); Tyr(SO3H); beta-Cyclohexyl-Ala;beta-(1-Cyclopentenyl)-Ala; beta-Cyclopentyl-Ala; beta-Cyclopropyl-Ala;beta-Quinolyl-Ala; beta-(2-Thiazolyl)-Ala; beta-(Triazole-1-yl)-Ala;beta-(2-Pyridyl)-Ala; beta-(3-Pyridyl)-Ala; Amino-Phe; Fluoro-Phe;Cyclohexyl-Gly; tBu-Gly; beta-(3-benzothienyl)-Ala;beta-(2-thienyl)-Ala; 5-Methyl-Trp; and A-Methyl-Trp. Proline residuescan be substituted with homopro (L-pipecolic acid); hydroxy-Pro;3,4-Dehydro-Pro; 4-fluoro-Pro; or alpha-methyl-Pro or anN(alpha)-C(alpha) cyclized amino acid analogues with the structure: n=0,1, 2, 3 Alanine residues can be substituted with alpha-substituted orN-methylated amino acid such as alpha-amino isobutyric acid (aib),L/D-alpha-ethylalanine (L/D-isovaline), L/D-methylvaline, orL/D-alpha-methylleucine or a non-natural amino acid such asbeta-fluoro-Ala. Alanine can also be substituted with: n=0, 1, 2, 3Glycine residues can be substituted with alpha-amino isobutyric acid(aib) or L/D-alpha-ethylalanine (L/D-isovaline).

Further examples of unnatural amino acids include: an unnatural analogof alanine (e.g., L-1-Nal or L-2-Nal); an unnatural analog of tyrosine;an unnatural analogue of glutamine; an unnatural analogue ofphenylalanine; an unnatural analogue of serine; an unnatural analogue ofthreonine; an alkyl, aryl, acyl, azido, cyano, halo, hydrazine,hydrazide, hydroxyl, alkenyl, alkynl, ether, thiol, sulfonyl, seleno,ester, thioacid, borate, boronate, phospho, phosphono, phosphine,heterocyclic, enone, imine, aldehyde, hydroxylamine, keto, or aminosubstituted amino acid, or any combination thereof; an amino acid with aphotoactivatable cross-linker; a spin-labeled amino acid; a fluorescentamino acid; an amino acid with a novel functional group; an amino acidthat covalently or noncovalently interacts with another molecule; ametal binding amino acid; an amino acid that is amidated at a site thatis not naturally amidated, a metal-containing amino acid; a radioactiveamino acid; a photocaged and/or photoisomerizable amino acid; a biotinor biotin-analogue containing amino acid; a glycosylated or carbohydratemodified amino acid; a keto containing amino acid; amino acidscomprising polyethylene glycol or polyether; a heavy atom substitutedamino acid (e.g., an amino acid containing deuterium, tritium, ¹³C, ¹⁵N,or ¹⁸O); a chemically cleavable or photocleavable amino acid; an aminoacid with an elongated side chain; an amino acid containing a toxicgroup; a sugar substituted amino acid, e.g., a sugar substituted serineor the like; a carbon-linked sugar-containing amino acid; a redox-activeamino acid; an α-hydroxy containing acid; an amino thio acid containingamino acid; an α, α disubstituted amino acid; a β-amino acid; a cyclicamino acid other than proline; an O-methyl-L-tyrosine; anL-3-(2-naphthyl)alanine; a 3-methyl-phenylalanine; aρ-acetyl-L-phenylalanine; an O-4-allyl-L-tyrosine; a4-propyl-L-tyrosine; a tri-O-acetyl-GlcNAc β-serine; an L-Dopa; afluorinated phenylalanine; an isopropyl-L-phenylalanine; ap-azido-L-phenylalanine; a p-acyl-L-phenylalanine; ap-benzoyl-L-phenylalanine; an L-phosphoserine; a phosphonoserine; aphosphonotyrosine; a p-iodo-phenylalanine; a 4-fluorophenylglycine; ap-bromophenylalanine; a p-amino-L-phenylalanine; anisopropyl-L-phenylalanine; L-3-(2-naphthyl)alanine;D-3-(2-naphthyl)alanine (dNal); an amino-, isopropyl-, orO-allyl-containing phenylalanine analogue; a dopa, O-methyl-L-tyrosine;a glycosylated amino acid; a p-(propargyloxy)phenylalanine;dimethyl-Lysine; hydroxy-proline; mercaptopropionic acid; methyl-lysine;3-nitro-tyrosine; norleucine; pyro-glutamic acid; Z (Carbobenzoxyl);ε-Acetyl-Lysine; β-alanine; β-aspartic acid; β-cyclohexylalanine;aminobenzoyl derivative; aminobutyric acid (Abu); citrulline;aminohexanoic acid (Ahx); aminoisobutyric acid (AIB); cyclohexylalanine;d-cyclohexylalanine; cyclohexylglycine; hydroxyproline; nitro-arginine;nitro-phenylalanine; nitro-tyrosine; norvaline; octahydroindolecarboxylate; ornithine (Orn); penicillamine (PEN);tetrahydroisoquinoline; diaminobutyric acid; diaminopimelic acid;pyroglutamic acid; homocysteine; homoserine; N-ε-dinitrophenyl-lysine;N-ε-methyl-lysine; N-ε-dimethyl-lysine; N,N,N-ε-trimethyl-lysine;acetamidomethyl protected amino acids and pegylated amino acids. Furtherexamples of unnatural amino acids and amino acid analogs can be found inU.S. 20030108885, U.S. 20030082575, US20060019347 (paragraphs 410-418)and the references cited therein. The polypeptides of the invention caninclude further modifications including those described inUS20060019347, paragraph 589.

“Nal” used herein refers to both L-1-naphthylalanine (L-1-Nal) andL-2-naphthylalanine (L-2-Nal).

In some embodiments, an amino acid can be replaced by anaturally-occurring, non-essential amino acid, e.g., taurine.

Alternatively, the Aad-GCRA peptides are cyclic peptides. Aad-GCRAcyclic peptides are prepared by methods known in the art. For example,macrocyclization is often accomplished by forming an amide bond betweenthe peptide N- and C-termini, between a side chain and the N- orC-terminus [e.g., with K₃Fe(CN)₆ at pH 8.5] (Samson et al.,Endocrinology, 137: 5182-5185 (1996)), or between two amino acid sidechains, such as cysteine. See, e.g., DeGrado, Adv Protein Chem, 39:51-124 (1988). In various aspects the Aad-GCRA peptides are [4,12; 7,15]bicycles.

In some Aad-GCRA peptides one or both members of one or both pairs ofCys residues which normally form a disulfide bond can be replaced byhomocysteine, penicillamine, 3-mercaptoproline (Kolodziej et al. 1996Int J Pept Protein Res 48:274); β, β dimethylcysteine (Hunt et al. 1993Int JPept Protein Res 42:249) or diaminopropionic acid (Smith et al.1978 J Med Chem 2 1:117) to form alternative internal cross-links at thepositions of the normal disulfide bonds.

In addition, one or more disulfide bonds can be replaced by alternativecovalent cross-links, e.g., an amide linkage (—CH2CH(O)NHCH 2- or—CH2NHCH(O)CH 2-), an ester linkage, a thioester linkage, a lactambridge (such as Asp [lactam]), a carbamoyl linkage, a urea linkage, athiourea linkage, a phosphonate ester linkage, an alkyl linkage(—CH2CH2CH2CH2-), an alkenyl linkage (—CH 2CH═CHCH 2-), an ether linkage(—CH2CH2OCH2- or —CH2OCH2CH2-), a thioether linkage (—CH2CH2SCH2- or—CH2SCH2CH2-), an amine linkage (—CH2CH2NHCH2- or —CH2NHCH 2CH2-) or athioamide linkage (—CH2CH(S)HNHCH 2- or —CH2NHCH(S)CH 2-). For example,Ledu et al. (Proc Nat'l Acad. Sci. 100:11263-78, 2003) describe methodsfor preparing lactam and amide cross-links.

The Aad-GCRA peptides can have one or more conventional polypeptidebonds replaced by an alternative bond. Such replacements can increasethe stability of the polypeptide. For example, replacement of thepolypeptide bond between a residue amino terminal to an aromatic residue(e.g. Tyr, Phe, Trp) with an alternative bond can reduce cleavage bycarboxy peptidases and may increase half-life in the digestive tract.Bonds that can replace polypeptide bonds include: a retro-inverso bond(C(O)—NH instead of NH—C(O); a reduced amide bond (NH—CH2); athiomethylene bond (S—CH2 or CH2-S); an oxomethylene bond (O—CH2 orCH2-O); an ethylene bond (CH2-CH2); a thioamide bond (C(S)—NH); atrans-olefine bond (CH═CH); a fluoro substituted trans-olefine bond(CF═CH); a ketomethylene bond (C(O)—CHR or CHR—C(O) wherein R is H orCH3; and a fluoro-ketomethylene bond (C(O)—CFR or CFR—C(O) wherein R isH or F or CH3.

The Aad-GCRA peptides can be modified using standard modifications.Modifications may occur at the amino (N-), carboxy (C-) terminus,internally or a combination of any of the preceeding. In one aspectdescribed herein, there may be more than one type of modification on thepolypeptide. Modifications include but are not limited to: acetylation,amidation, biotinylation, cinnamoylation, farnesylation, formylation,myristoylation, palmitoylation, phosphorylation (Ser, Tyr or Thr),stearoylation, succinylation, sulfurylation and cyclisation (viadisulfide bridges or amide cyclisation), and modification by Cys3 orCys5. The Aad-GCRA peptides described herein may also be modified by 2,4-dinitrophenyl (DNP), DNP-lysine, modification by7-Amino-4-methyl-coumarin (AMC), flourescein, NBD(7-Nitrobenz-2-Oxa-1,3-Diazole), p-nitro-anilide, rhodamine B, EDANS(5-((2-aminoethyl)amino)naphthalene-1-sulfonic acid), dabcyl, dabsyl,dansyl, texas red, FMOC, and Tamra (Tetramethylrhodamine). The Aad-GCRApeptides described herein may also be conjugated to, for example,polyethylene glycol (PEG); alkyl groups (e.g., C1-C20 straight orbranched alkyl groups); fatty acid radicals; combinations of PEG, alkylgroups and fatty acid radicals (See, U.S. Pat. No. 6,309,633; Soltero etal., 2001 Innovations in Pharmaceutical Technology 106-110); BSA and KLH(Keyhole Limpet Hemocyanin). The addition of PEG and other polymerswhich can be used to modify polypeptides of the invention is describedin US2006019347 section IX.

Also included in the invention are peptides that biologically orfunctional equivalent to the peptides described herein. The term“biologically equivalent” or functional equivalent” is intended to meanthat the compositions of the present invention are capable ofdemonstrating some or all of the cGMP production modulatory effects.

Aad-GCRA peptides can also include derivatives of Aad-GCRA peptideswhich are intended to include hybrid and modified forms of Aad-GCRApeptides in which certain amino acids have been deleted or replaced andmodifications such as where one or more amino acids have been changed toa modified amino acid or unusual amino acid and modifications such asglycosylation so long the modified form retains the biological activityof Aad-GCRA peptides. By retaining the biological activity, it is meantthat cGMP and or apoptosis is induced by the Aad-GCRA peptide, althoughnot necessarily at the same level of potency as that of anaturally-occurring GCRA peptide identified.

Preferred variants are those that have conservative amino acidsubstitutions made at one or more predicted non-essential amino acidresidues. A “conservative amino acid substitution” is one in which theamino acid residue is replaced with an amino acid residue having asimilar side chain. Families of amino acid residues having similar sidechains have been defined in the art. These families include amino acidswith basic side chains (e.g., lysine, arginine, histidine), acidic sidechains (e.g., aspartic acid, glutamic acid), uncharged polar side chains(e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine,cysteine), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine, tryptophan),beta-branched side chains (e.g., threonine, valine, isoleucine) andaromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,histidine). Thus, a predicted nonessential amino acid residue in anAad-GCRA polypeptide is replaced with another amino acid residue fromthe same side chain family. Alternatively, in another embodiment,mutations can be introduced randomly along all or part of an Aad-GCRAcoding sequence, such as by saturation mutagenesis, and the resultantmutants can be screened to identify mutants that retain activity.

Also included within the meaning of substantially homologous is anyAad-GCRA peptide which may be isolated by virtue of cross-reactivitywith antibodies to the Aad-GCRA peptide.

Preparation of Aad-GCRA Peptides

Aad-GCRA peptides are easily prepared using modern cloning techniques,or may be synthesized by solid state methods or by site-directedmutagenesis. An Aad-GCRA peptide may include dominant negative forms ofa polypeptide.

Chemical synthesis may generally be performed using standard solutionphase or solid phase peptide synthesis techniques, in which a peptidelinkage occurs through the direct condensation of the amino group of oneamino acid with the carboxy group of the other amino acid with theelimination of a water molecule. Peptide bond synthesis by directcondensation, as formulated above, requires suppression of the reactivecharacter of the amino group of the first and of the carboxyl group ofthe second amino acid. The masking substituents must permit their readyremoval, without inducing breakdown of the labile peptide molecule.

In solution phase synthesis, a wide variety of coupling methods andprotecting groups may be used (See, Gross and Meienhofer, eds., “ThePeptides: Analysis, Synthesis, Biology,” Vol. 1-4 (Academic Press,1979); Bodansky and Bodansky, “The Practice of Peptide Synthesis,” 2ded. (Springer Verlag, 1994)). In addition, intermediate purification andlinear scale up are possible. Those of ordinary skill in the art willappreciate that solution synthesis requires consideration of main chainand side chain protecting groups and activation method. In addition,careful segment selection is necessary to minimize racemization duringsegment condensation. Solubility considerations are also a factor. Solidphase peptide synthesis uses an insoluble polymer for support duringorganic synthesis. The polymer-supported peptide chain permits the useof simple washing and filtration steps instead of laboriouspurifications at intermediate steps. Solid-phase peptide synthesis maygenerally be performed according to the method of Merrifield et al., J.Am. Chem. Soc., 1963, 85:2149, which involves assembling a linearpeptide chain on a resin support using protected amino acids. Solidphase peptide synthesis typically utilizes either the Boc or Fmocstrategy, which is well known in the art.

Those of ordinary skill in the art will recognize that, in solid phasesynthesis, deprotection and coupling reactions must go to completion andthe side-chain blocking groups must be stable throughout the synthesis.In addition, solid phase synthesis is generally most suitable whenpeptides are to be made on a small scale.

Acetylation of the N-terminal can be accomplished by reacting the finalpeptide with acetic anhydride before cleavage from the resin.C-amidation is accomplished using an appropriate resin such asmethylbenzhydrylamine resin using the Boc technology.

Alternatively the Aad-GCRA peptides are produced by modern cloningtechniques. For example, the Aad-GCRA peptides are produced either inbacteria including, without limitation, E. coli, or in other existingsystems for polypeptide or protein production (e.g., Bacillus subtilis,baculovirus expression systems using Drosophila Sf9 cells, yeast orfilamentous fungal expression systems, mammalian cell expressionsystems), or they can be chemically synthesized. If the Aad-GCRA peptideor variant peptide is to be produced in bacteria, e.g., E. coli, thenucleic acid molecule encoding the polypeptide may also encode a leadersequence that permits the secretion of the mature polypeptide from thecell. Thus, the sequence encoding the polypeptide can include the presequence and the pro sequence of, for example, a naturally-occurringbacterial ST polypeptide. The secreted, mature polypeptide can bepurified from the culture medium.

The sequence encoding an Aad-GCRA peptide described herein can beinserted into a vector capable of delivering and maintaining the nucleicacid molecule in a bacterial cell. The DNA molecule may be inserted intoan autonomously replicating vector (suitable vectors include, forexample, pGEM3Z and pcDNA3, and derivatives thereof). The vector nucleicacid may be a bacterial or bacteriophage DNA such as bacteriophagelambda or M13 and derivatives thereof. Construction of a vectorcontaining a nucleic acid described herein can be followed bytransformation of a host cell such as a bacterium. Suitable bacterialhosts include but are not limited to, E. coli, B subtilis, Pseudomonas,Salmonella. The genetic construct also includes, in addition to theencoding nucleic acid molecule, elements that allow expression, such asa promoter and regulatory sequences. The expression vectors may containtranscriptional control sequences that control transcriptionalinitiation, such as promoter, enhancer, operator, and repressorsequences.

A variety of transcriptional control sequences are well known to thosein the art. The expression vector can also include a translationregulatory sequence (e.g., an untranslated 5′ sequence, an untranslated3′ sequence, or an internal ribosome entry site). The vector can becapable of autonomous replication or it can integrate into host DNA toensure stability during polypeptide production.

The protein coding sequence that includes an Aad-GCRA peptide describedherein can also be fused to a nucleic acid encoding a polypeptideaffinity tag, e.g., glutathione S-transferase (GST), maltose E bindingprotein, protein A, FLAG tag, hexa-histidine, myc tag or the influenzaHA tag, in order to facilitate purification. The affinity tag orreporter fusion joins the reading frame of the polypeptide of interestto the reading frame of the gene encoding the affinity tag such that atranslational fusion is generated. Expression of the fusion gene resultsin translation of a single polypeptide that includes both thepolypeptide of interest and the affinity tag. In some instances whereaffinity tags are utilized, DNA sequence encoding a protease recognitionsite will be fused between the reading frames for the affinity tag andthe polypeptide of interest.

Genetic constructs and methods suitable for production of immature andmature forms of the Aad-GCRA peptides and variants described herein inprotein expression systems other than bacteria, and well known to thoseskilled in the art, can also be used to produce polypeptides in abiological system.

The peptides disclosed herein may be modified by attachment of a secondmolecule that confers a desired property upon the peptide, such asincreased half-life in the body, for example, pegylation. Suchmodifications also fall within the scope of the term “variant” as usedherein.

Compositions

The present invention also provides compositions comprising at least oneAad-GCRA peptide, at least one enteric coating which releases thepeptide at a specific pH (e.g., about pH 4.0, pH 5.0, pH 6.0 or pH 7.0)and an inert carrier.

A composition may comprise an enteric coating which releases the peptideat pH5 and an inert carrier coated with Aad-GCRA peptides.

A composition may comprise an enteric coating which releases the peptideat pH6 and an inert carrier coated with Aad-GCRA peptides.

A composition may comprise an enteric coating which releases the peptideat pH7 and an inert carrier coated with Aad-GCRA peptides.

The present invention further provides a formulation comprising amixture of compositions that contain different peptides and/or thatrelease the peptides at different pH levels. The mixture may comprise atleast 2, 3, 4 or more compositions that release the peptides atdifferent pH levels. The mixture may comprise at least 2, 3, 4 or morecompositions that contain different Aad-GCRA peptides. A skilled artisancan determine the ratio of these compositions within the mixture, forexample, according to the activity of each peptide, solubility of eachpeptide, and/or the targeting region of the GI tract.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with Aad-GCRA peptides and anenteric coating that releases the peptides at pH5.0 (“pH5.0composition”) and (2) a composition having an inert carrier coated withAad-GCRA peptides and an enteric coating that releases the peptides atpH6.0 (“pH6.0 composition”).

The ratio of pH5.0 composition to pH6.0 composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofpH5.0 composition to pH6.0 composition is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with Aad-GCRA peptides and anenteric coating that releases the peptides at pH5.0 (“pH5.0composition”) and (2) a composition having an inert carrier coated withAad-GCRA peptides and an enteric coating that releases the peptides atpH7.0 (“pH7.0 composition”).

The ratio of pH5.0 composition to pH7.0 composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofpH5.0 composition to pH7.0 composition is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with Aad-GCRA peptides and anenteric coating that releases the peptides at pH6.0 (“pH6.0composition”) and (2) a composition having an inert carrier coated withAad-GCRA peptides and an enteric coating that releases the peptides atpH7.0 (“pH7.0 composition”).

The ratio of pH6.0 composition to pH7.0 composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofpH6.0 composition to pH7.0 composition is 10:1, 9:1, 8:1, 7:1, 6:1, 5:1,4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with Aad-GCRA peptides and anenteric coating that releases the peptides at pH5.0 (“pH5.0composition”); (2) a composition having an inert carrier coated withAad-GCRA peptides and an enteric coating that releases the peptides atpH6.0 (“pH6.0 composition”) and (3) a composition having an inertcarrier coated with Aad-GCRA and an enteric coating that releases thepeptides at pH7.0 (“pH7.0 composition”).

The ratio of pH5.0 composition to pH6.0 composition to pH7.0 compositioncan be determined, for example, by the activity of each peptide,solubility of each peptide, and/or the targeting region of the GI tract.

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with Aad-GCRA peptides and anenteric coating that releases the peptides at duodenum or jejunum(“duodenum composition”) and (2) a composition having an inert carriercoated with Aad-GCRA peptides and an enteric coating that releases thepeptides at ileum, terminal ileum, or ascending colon (“ileumcomposition”).

In some embodiments, a formulation comprises a mixture of (1) acomposition having an inert carrier coated with Aad-GCRA peptides and anenteric coating that releases the peptides in a pH range of 4.5 to 5.5or in a pH range of 5.5 to 6.5 at duodenum or jejunum (“duodenumcomposition”); and (2) a composition having an inert carrier coated withAad-GCRA peptides and an enteric coating that releases the peptides in apH range of 5.5 to 6.5 or in a pH range of 6.5 to 7.5 at ileum, terminalileum, or ascending colon (“ileum composition”).

The ratio of duodenum composition to ileum composition can be any valuebetween 100:1 (v/v) and 1:100 (v/v) and can be determined, for example,by the activity of each peptide, solubility of each peptide, and/or thetargeting region of the GI tract. In some embodiments, the ratio ofduodenum composition to ileum composition is 10:1, 9:1, 8:1, 7:1, 6:1,5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or1:10.

The targeting region of the GI track includes, but is not limited to,duodenum, jejunum, ileum, terminal ileum, and ascending colon.

In some embodiments, the inert carrier is selected from the groupconsisting of sorbitol, mannitol, EMDEX, and starch. In someembodiments, the carrier is mannitol (e.g., MANNOGEM) ormicrocrystalline cellulose (e.g., PROSOLV, CELPHERE®, CELPHERE® beads).In a preferred embodiment, the carrier is microcrystalline cellulosespheres or spherical microcrystalline cellulose, such as Celphere®SCP-100.

The enteric coating material is chosen to target the release of thecomposition of the present invention to a specific region of thegastrointestinal tract. The enteric coating material preferablycomprises one of the following: (1) a pH dependent polymer; (2) aswellable polymer; or (3) a degradable composition. More coatingmaterials and formulations can be found in PCT publications WO10/065751, WO 12/118972, and WO 12/037380 and US publication20120237593, each of which is incorporated herein by reference in itsentirety.

Therapeutic Methods

The present invention provides for both prophylactic and therapeuticmethods of treating a subject at risk of (or susceptible to) developinga disorder or having a disorder that is mediated by guanylate cyclasereceptor agonists by administering an Aad-GCRA peptide described herein.

The present invention also provides methods for treating a conditionthat responds to enhanced cGMP levels in a subject by administering anAad-GCRA peptide described herein.

Disorders mediated by the guanylate cyclase receptor agonists andconditions that respond to enhanced cGMP levels include lipid metabolismdisorders, biliary disorders, gastrointestinal disorders, inflammatorydisorders, lung disorders, cancer, cardiac disorders includingcardiovascular disorders, eye disorders, oral disorders, blooddisorders, liver disorders, skin disorders, prostate disorders,endocrine disorders, increasing gastrointestinal motility and obesity.Lipid metabolism disorders include, but not limited to, dyslipidemia,hyperlipidemia, hypercholesterolemia, hypertriglyceridemia,sitosterolemia, familial hypercholesterolemia, xanthoma, combinedhyperlipidemia, lecithin cholesterol acyltransferase deficiency, tangierdisease, abetalipoproteinemia, erectile dysfunction, fatty liverdisease, and hepatitis. Billary disorders include gallbladder disorderssuch as for example, gallstones, gall bladder cancer cholangitis, orprimary sclerosing cholangitis; or bile duct disorders such as forexample, cholecystitis, bile duct cancer or fascioliasis.Gastointestinal disorders include for example, inflammatory boweldisease (IBD), irritable bowel syndrome (IBS), non-ulcer dyspepsia,chronic intestinal pseudo-obstruction, functional dyspepsia, colonicpseudo-obstruction, duodenogastric reflux, gastroesophageal refluxdisease (GERD), ileus inflammation (e.g., post-operative ileus),gastroparesis, heartburn (high acidity in the GI tract), constipation(e.g., chronic constipation, constipation associated with IBS,constipation associated with use of medications such as opioids,osteoarthritis drugs, osteoporosis drugs; post surgical constipation,constipation associated with neuropathic disorders). Inflammatorydisorders include tissue and organ inflammation such as kidneyinflammation (e.g., nephritis), gastrointestinal system inflammation(e.g., Crohn's disease and ulcerative colitis), IBD, necrotizingenterocolitis (NEC); pancreatic inflammation (e.g., pancreatis), lunginflammation (e.g., bronchitis or asthma) or skin inflammation (e.g.,psoriasis, eczema). Lung Disorders include for example chronicobstructive pulmonary disease (COPD), and fibrosis. Cancer includestissue and organ carcinogenesis including metatases such as for examplegastrointestinal cancer (e.g., gastric cancer, esophageal cancer,pancreatic cancer colorectal cancer, intestinal cancer, anal cancer,liver cancer, gallbladder cancer, or colon cancer); lung cancer; thyroidcancer; skin cancer (e.g., melanoma); oral cancer; urinary tract cancer(e.g. bladder cancer or kidney cancer); blood cancer (e.g. myeloma orleukemia) or prostate cancer. Cardiac disorders include for example,congestive heart failure, trachea cardia hypertension, high cholesterol,or high tryglycerides. Cardiovascular disorders include for exampleaneurysm, angina, atherosclerosis, cerebrovascular accident (stroke),cerebrovasculardisease, congestive heart failure, coronary arterydisease, myocardial infarction (heart attack), or peripheral vasculardisease. Liver disorders include for example cirrhosis and fibrosis. Inaddition, Aad derivatives of GC-C agonist described herein may also beuseful to facilitate liver regeneration in liver transplant patients.Eye disorders include for example increased intra-ocular pressure,glaucoma, dry eyes retinal degeneration, disorders of tear glands or eyeinflammation. Skin disorders include for example xerosis. Oral disordersinclude for example dry mouth (xerostomia), Sjögren's syndrome, gumdiseases (e.g., periodontal disease), or salivary gland duct blockage ormalfunction. Prostate disorders include for example benign prostatichyperplasia (BPH). Endocrine disorders include for example diabetesmellitus, hyperthyroidism, hypothyroidism, and cystic fibrosis.

The term “treatment” refers to reducing or alleviating symptoms in asubject, preventing symptoms from worsening or progressing, and/orpreventing disease in a subject who is free therefrom. For a givensubject, improvement in a symptom, its worsening, regression, orprogression may be determined by any objective or subjective measure.Efficacy of the treatment may be measured as an improvement in morbidityor mortality (e.g., lengthening of survival curve for a selectedpopulation). Thus, effective treatment would include therapy of existingdisease, control of disease by slowing or stopping its progression,prevention of disease occurrence, reduction in the number or severity ofsymptoms, or a combination thereof. The effect may be shown in acontrolled study using one or more statistically significant criteria.

Intracellular cGMP is produced by exposing, e.g., contacting a tissue(e.g., gastrointestinals tissue) or cell with Aad-GCRA peptides. Byinducing is meant an increase in cGMP production compared to a tissue orcell that has not been in contact with Aad-GCRA peptide or variant.Tissues or cells are directly contacted with an Aad-GCRA peptide orvariant. Alternatively, the Aad-GCRA peptide or variant is administeredsystemically. Aad-GCRA peptides or variants are administered in anamount sufficient to increase intracellular cGMP concentration. cGMPproduction is measured by a cell-based assay known in the art (25).

Disorders are treated, prevented or alleviated by administering to asubject, e.g., a mammal such as a human in need thereof, atherapeutically effective dose of any Aad-GCRA peptide described herein.

The Aad-GCRA peptides may be in a pharmaceutical composition in unitdose form, together with one or more pharmaceutically acceptableexcipients. The term “unit dose form” refers to a single drug deliveryentity, e.g., a tablet, capsule, solution or inhalation formulation. Theamount of peptide present should be sufficient to have a positivetherapeutic effect when administered to a patient (typically, between 10μg and 3 g). What constitutes a “positive therapeutic effect” willdepend upon the particular condition being treated and will include anysignificant improvement in a condition readily recognized by one ofskill in the art.

The present invention also provides a method of colonic cleansing byadministering to a subject in need thereof an effective amount of anycompositions of the present invention, for example an Aad-GCRA peptidedescribed herein.

This method can be used in cleansing or purging the bowels or colonprior to carrying out a diagnostic, therapeutic or surgical procedure onthe colon, rectum or anus or elsewhere in the abdomen. The diagnostic orsurgical procedure may, for example, be sigmoidoscopy, colonoscopy,radiographic examination, preparation for patients undergoing bowelsurgery, and other medical or diagnostic procedures. It has beenbelieved that profuse, uncontrolled diarrhea was necessary to produceadequate cleansing of the colon. This present invention provides a safeand effective cleansing method for the bowels and colon, without theingestion of large volumes of lavage solutions, without the unpleasant,bitter, and dangerous hypertonic salt solutions, thus providing animproved patients compliance.

“Subject”, as used herein, means an individual. In one aspect, thesubject is a mammal such as a primate, and, in another aspect, thesubject is a human. The term “subject” also includes domesticatedanimals (e.g., cats, dogs, etc.), and livestock (e.g., cattle, horses,pigs, sheep, goats, etc.). The subject may be at risk of (or susceptibleto) developing a disorder that is mediated by guanylate cyclase receptoragonists or may have a disorder that is mediated by guanylate cyclasereceptor agonists. The subject may be a human over 50 years old.

Preferably, the Aad-GCRA peptide used for any methods described hereinisAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶(SEQ ID NO: 1),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶ (SEQ ID NO: 32),Asn¹-Asp²-Aad³-Cys⁴-Glu-Leu⁶-Cys⁷-Val¹⁰-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Thr¹⁶(SEQ ID NO: 119),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶(SEQ ID NO: 120),dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶(SEQ ID NO: 17), orpyGlu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶(SEQ ID NO: 56).

The Aad-GCRA peptides can be administered alone or in combination withother agents. For example the Aad-GCRA peptides can be administered incombination with inhibitors of cGMP dependent phosphodiesterase, suchas, for example, sulindac sulfone, zaprinast, motapizone, vardenafil,and sildenafil; one or more other chemotherapeutic agents; oranti-inflammatory drugs such as, for example, steroids or non-steroidalanti-inflammatory drugs (NSAIDs), such as aspirin.

The Aad-GCRA peptides described herein can be administered with one ormore other agents, for example, L-glucose, cholera toxin, osmoticcolonic evacuants, cathartic, laxatives and agents for treating chronicconstipation, or any combination thereof, for cleansing or purging thebowels or colon prior to carrying out a diagnostic, therapeutic orsurgical procedure on the colon, rectum or anus or elsewhere in theabdomen. In some embodiments, the compositions (e.g., Aad-GCRA peptides)or the formulations described herein can be administered with L-glucose.

Combination therapy can be achieved by administering two or more agents,e.g., an Aad-GCRA peptide described herein and another compound, each ofwhich is formulated and administered separately, or by administering twoor more agents in a single formulation. Other combinations are alsoencompassed by combination therapy. For example, two agents can beformulated together and administered in conjunction with a separateformulation containing a third agent. While the two or more agents inthe combination therapy can be administered simultaneously, they neednot be. For example, administration of a first agent (or combination ofagents) can precede administration of a second agent (or combination ofagents) by minutes, hours, days, or weeks. Thus, the two or more agentscan be administered within minutes of each other or within 1, 2, 3, 6,9, 12, 15, 18, or 24 hours of each other or within 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 12, 14 days of each other or within 2, 3, 4, 5, 6, 7, 8, 9, or10 weeks of each other. In some cases even longer intervals arepossible. While in many cases it is desirable that the two or moreagents used in a combination therapy be present in within the patient'sbody at the same time, this need not be so.

The Aad-GCRA peptides described herein may be combined withcGMP-dependent phosphodiesterase inhibitors, e.g., sulindac sulfone,zaprinast, motapizone, vardenafil, and sildenafil to further enhancelevels of cGMP in the target tissues or organs.

Combination therapy can also include two or more administrations of oneor more of the agents used in the combination. For example, if agent Xand agent Y are used in a combination, one could administer themsequentially in any combination one or more times, e.g., in the orderX-Y-X, X-X-Y, Y-X-Y, Y-Y-X, X-X-Y-Y, etc.

Combination therapy can also include the administration of one of theAad-GCRA peptides with azothioprine and/or other immunomodulatingagents. The immunomodulating agents may include small molecule drugs andbiologics such as Remicade, Humaira, Cimzia etc.

Combination therapy can also include the administration of two or moreagents via different routes or locations. For example, (a) one agent isadministered orally and another agent is administered intravenously or(b) one agent is administered orally and another is administeredlocally. In each case, the agents can either simultaneously orsequentially. Approximate dosages for some of the combination therapyagents described herein are found in the “BNF Recommended Dose” columnof tables on pages 11-17 of WO01/76632 (the data in the tables beingattributed to the March 2000 British National Formulary) and can also befound in other standard formularies and other drug prescribingdirectories. For some drugs, the customary prescribed dose for anindication will vary somewhat from country to country.

The Aad-GCRA peptides, alone or in combination, can be combined with anypharmaceutically acceptable carrier or medium. Thus, they can becombined with materials that do not produce an adverse, allergic orotherwise unwanted reaction when administered to a patient. The carriersor mediums used can include solvents, dispersants, coatings, absorptionpromoting agents, controlled release agents, and one or more inertexcipients (which include starches, polyols, granulating agents,microcrystalline cellulose (e.g. celphere, Celphere Beads®), diluents,lubricants, binders, disintegrating agents, and the like), etc. Ifdesired, tablet dosages of the disclosed compositions may be coated bystandard aqueous or nonaqueous techniques.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical),transmucosal, and rectal administration. Solutions or suspensions usedfor parenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid;buffers such as acetates, citrates or phosphates, and agents for theadjustment of tonicity such as sodium chloride or dextrose. The pH canbe adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringeability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol, sorbitol, sodium chloride in thecomposition. Prolonged absorption of the injectable compositions can bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound (e.g., an Aad-GCRA peptide) in the required amount in anappropriate solvent with one or a combination of ingredients enumeratedabove, as required, followed by filtered sterilization. Generally,dispersions are prepared by incorporating the active compound into asterile vehicle that contains a basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, methods ofpreparation are vacuum drying and freeze-drying that yields a powder ofthe active ingredient plus any additional desired ingredient from apreviously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an ediblecarrier. Such as mannitol, fructooligosaccharides, polyethylene glycoland other excipients. They can be enclosed in gelatin capsules orcompressed into tablets. For the purpose of oral therapeuticadministration, the active compound can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Oral compositionscan also be prepared using a fluid carrier for use as a mouthwash,wherein the compound in the fluid carrier is applied orally and swishedand expectorated or swallowed. Pharmaceutically compatible bindingagents, and/or adjuvant materials can be included as part of thecomposition. The tablets, pills, capsules, troches and the like cancontain any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate or Sterotes; a glidant such as colloidal silicondioxide; a sweetening agent such as sucrose or saccharin; or a flavoringagent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives.

Transmucosal administration can be accomplished through the use of nasalsprays or suppositories. For transdermal administration, the activecompounds are formulated into ointments, salves, gels, or creams asgenerally known in the art.

The compounds can also be prepared in the form of suppositories (e.g.,with conventional suppository bases such as cocoa butter and otherglycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811, incorporated fully herein by reference.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

Compositions of the present invention may also optionally include othertherapeutic ingredients, anti-caking agents, preservatives, sweeteningagents, colorants, flavors, desiccants, plasticizers, dyes, glidants,anti-adherents, anti-static agents, surfactants (wetting agents),antioxidants, film-coating agents, and the like. Any such optionalingredient must be compatible with the compound described herein toinsure the stability of the formulation.

The composition may contain other additives as needed, including forexample lactose, glucose, fructose, galactose, trehalose, sucrose,maltose, raffnose, maltitol, melezitose, stachyose, lactitol,palatinite, starch, xylitol, mannitol, myoinositol, and the like, andhydrates thereof, and amino acids, for example alanine, glycine andbetaine, and polypeptides and proteins, for example albumen.

Examples of excipients for use as the pharmaceutically acceptablecarriers and the pharmaceutically acceptable inert carriers and theaforementioned additional ingredients include, but are not limited tobinders, fillers, disintegrants, lubricants, anti-microbial agents, andcoating agents such as: BINDERS: corn starch, potato starch, otherstarches, gelatin, natural and synthetic gums such as acacia, xanthan,sodium alginate, alginic acid, other alginates, powdered tragacanth,guar gum, cellulose and its derivatives (e.g., ethyl cellulose,cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethylcellulose), polyvinyl pyrrolidone (e.g., povidone, crospovidone,copovidone, etc), methyl cellulose, Methocel, pre-gelatinized starch(e.g., STARCH 1500® and STARCH 1500 LM®, sold by Colorcon, Ltd.),hydroxypropyl methyl cellulose, microcrystalline cellulose (FMCCorporation, Marcus Hook, Pa., USA), or mixtures thereof, FILLERS: talc,calcium carbonate (e.g., granules or powder), dibasic calcium phosphate,tribasic calcium phosphate, calcium sulfate (e.g., granules or powder),microcrystalline cellulose, powdered cellulose, dextrates, kaolin,mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch,dextrose, fructose, honey, lactose anhydrate, lactose monohydrate,lactose and aspartame, lactose and cellulose, lactose andmicrocrystalline cellulose, maltodextrin, maltose, mannitol,microcrystalline cellulose &amp; guar gum, molasses, sucrose, ormixtures thereof, DISINTEGRANTS: agar-agar, alginic acid, calciumcarbonate, microcrystalline cellulose, croscarmellose sodium,crospovidone, polacrilin potassium, sodium starch glycolate, potato ortapioca starch, other starches, pre-gelatinized starch, clays, otheralgins, other celluloses, gums (like gellan), low-substitutedhydroxypropyl cellulose, or mixtures thereof, LUBRICANTS: calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, sodium stearyl fumarate, vegetable based fattyacids lubricant, talc, hydrogenated vegetable oil (e.g., peanut oil,cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil andsoybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, syloidsilica gel (AEROSIL 200, W.R. Grace Co., Baltimore, Md. USA), acoagulated aerosol of synthetic silica (Deaussa Co., Piano, Tex. USA), apyrogenic silicon dioxide (CAB-O-SIL, Cabot Co., Boston, Mass. USA), ormixtures thereof, ANTI-CAKING AGENTS: calcium silicate, magnesiumsilicate, silicon dioxide, colloidal silicon dioxide, talc, or mixturesthereof, ANTIMICROBIAL AGENTS: benzalkonium chloride, benzethoniumchloride, benzoic acid, benzyl alcohol, butyl paraben, cetylpyridiniumchloride, cresol, chlorobutanol, dehydroacetic acid, ethylparaben,methylparaben, phenol, phenylethyl alcohol, phenoxyethanol,phenylmercuric acetate, phenylmercuric nitrate, potassium sorbate,propylparaben, sodium benzoate, sodium dehydroacetate, sodiumpropionate, sorbic acid, thimersol, thymo, or mixtures thereof, andCOATING AGENTS: sodium carboxymethyl cellulose, cellulose acetatephthalate, ethylcellulose, gelatin, pharmaceutical glaze, hydroxypropylcellulose, hydroxypropyl methylcellulose (hypromellose), hydroxypropylmethyl cellulose phthalate, methylcellulose, polyethylene glycol,polyvinyl acetate phthalate, shellac, sucrose, titanium dioxide,carnauba wax, microcrystalline wax, gellan gum, maltodextrin,methacrylates, microcrystalline cellulose and carrageenan or mixturesthereof.

The formulation can also include other excipients and categories thereofincluding but not limited to L-histidine, Pluronic®, Poloxamers (such asLutrol® and Poloxamer 188), ascorbic acid, glutathione, permeabilityenhancers (e.g. lipids, sodium cholate, acylcarnitine, salicylates,mixed bile salts, fatty acid micelles, chelators, fatty acid,surfactants, medium chain glycerides), protease inhibitors (e.g. soybeantrypsin inhibitor, organic acids), pH lowering agents and absorptionenhancers effective to promote bioavailability (including but notlimited to those described in U.S. Pat. No. 6,086,918 and U.S. Pat. No.5,912,014), creams and lotions (like maltodextrin and carrageenans);materials for chewable tablets (like dextrose, fructose, lactosemonohydrate, lactose and aspartame, lactose and cellulose, maltodextrin,maltose, mannitol, microcrystalline cellulose and guar gum, sorbitolcrystalline); parenterals (like mannitol and povidone); plasticizers(like dibutyl sebacate, plasticizers for coatings, polyvinylacetatephthalate); powder lubricants (like glyceryl behenate); soft gelatincapsules (like sorbitol special solution); spheres for coating (likesugar spheres); spheronization agents (like glyceryl behenate andmicrocrystalline cellulose); suspending/gelling agents (likecarrageenan, gellan gum, mannitol, microcrystalline cellulose, povidone,sodium starch glycolate, xanthan gum); sweeteners (like aspartame,aspartame and lactose, dextrose, fructose, honey, maltodextrin, maltose,mannitol, molasses, sorbitol crystalline, sorbitol special solution,sucrose); wet granulation agents (like calcium carbonate, lactoseanhydrous, lactose monohydrate, maltodextrin, mannitol, microcrystallinecellulose, povidone, starch), caramel, carboxymethylcellulose sodium,cherry cream flavor and cherry flavor, citric acid anhydrous, citricacid, confectioner's sugar, D&C Red No. 33, D&C Yellow #10 AluminumLake, disodium edetate, ethyl alcohol 15%, FD&C Yellow No. 6 aluminumlake, FD&C Blue #1 Aluminum Lake, FD&C Blue No. 1, FD&C blue no. 2aluminum lake, FD&C Green No. 3, FD&C Red No. 40, FD&C Yellow No. 6Aluminum Lake, FD&C Yellow No. 6, FD&C Yellow No. 10, glycerolpalmitostearate, glyceryl monostearate, indigo carmine, lecithin,manitol, methyl and propyl parabens, mono ammonium glycyrrhizinate,natural and artificial orange flavor, pharmaceutical glaze, poloxamer188, Polydextrose, polysorbate 20, polysorbate 80, polyvidone,pregelatinized corn starch, pregelatinized starch, red iron oxide,saccharin sodium, sodium carboxymethyl ether, sodium chloride, sodiumcitrate, sodium phosphate, strawberry flavor, synthetic black ironoxide, synthetic red iron oxide, titanium dioxide, and white wax.

Solid oral dosage forms may optionally be treated with coating systems(e.g. Opadry® fx film coating system, for example Opadry® blue(OY-LS-20921), Opadry® white (YS-2-7063), Opadry® white (YS-1-7040), andblack ink (S-1-8 106).

The agents either in their free form or as a salt can be combined with apolymer such as polylactic-glycoloic acid (PLGA),poly-(I)-lactic-glycolic-tartaric acid (P(I)LGT) (WO 01/12233),polyglycolic acid (U.S. Pat. No. 3,773,919), polylactic acid (U.S. Pat.No. 4,767,628), poly(ε-caprolactone) and poly(alkylene oxide) (U.S.20030068384) to create a sustained release formulation. Suchformulations can be used to implants that release a polypeptide oranother agent over a period of a few days, a few weeks or several monthsdepending on the polymer, the particle size of the polymer, and the sizeof the implant (See, e.g., U.S. Pat. No. 6,620,422). Other sustainedrelease formulations and polymers for use in are described in EP 0 467389 A2, WO 93/24150, U.S. Pat. No. 5,612,052, WO 97/40085, WO 03/075887,WO 01/01964A2, U.S. Pat. No. 5,922,356, WO 94/155587, WO 02/074247A2, WO98/25642, U.S. Pat. No. 5,968,895, U.S. Pat. No. 6,180,608, U.S.20030171296, U.S. 20020176841, U.S. Pat. No. 5,672,659, U.S. Pat. No.5,893,985, U.S. Pat. No. 5,134,122, U.S. Pat. No. 5,192,741, U.S. Pat.No. 5,192,741, U.S. Pat. No. 4,668,506, U.S. Pat. No. 4,713,244, U.S.Pat. No. 5,445,832 U.S. Pat. No. 4,931,279, U.S. Pat. No. 5,980,945, WO02/058672, WO 97/26015, WO 97/04744, and US20020019446. In suchsustained release formulations microparticles (Delie and Blanco-Prieto2005 Molecule 10:65-80) of polypeptide are combined with microparticlesof polymer. One or more sustained release implants can be placed in thelarge intestine, the small intestine or both. U.S. Pat. No. 6,011,011and WO 94/06452 describe a sustained release formulation providingeither polyethylene glycols (i.e. PEG 300 and PEG 400) or triacetin. WO03/053401 describes a formulation which may both enhance bioavailabilityand provide controlled release of the agent within the GI tract.Additional controlled release formulations are described in WO 02/38129,EP 326151, U.S. Pat. No. 5,236,704, WO 02/30398, WO 98/13029; U.S.20030064105, U.S. 20030138488A1, U.S. 20030216307A1, U.S. Pat. No.6,667,060, WO 01/49249, WO 01/49311, WO 01/49249, WO 01/49311, and U.S.Pat. No. 5,877,224 materials which may include those described inWO04041195 (including the seal and enteric coating described therein)and pH-sensitive coatings that achieve delivery in the colon includingthose described in U.S. Pat. No. 4,910,021 and WO9001329. U.S. Pat. No.4,910,021 describes using a pH-sensitive material to coat a capsule.WO9001329 describes using pH-sensitive coatings on beads containingacid, where the acid in the bead core prolongs dissolution of thepH-sensitive coating. U.S. Pat. No. 5,175,003 discloses a dual mechanismpolymer mixture composed of pH-sensitive enteric materials andfilm-forming plasticizers capable of conferring permeability to theenteric material, for use in drug-delivery systems; a matrix pelletcomposed of a dual mechanism polymer mixture permeated with a drug andsometimes covering a pharmaceutically neutral nucleus; a membrane-coatedpellet comprising a matrix pellet coated with a dual mechanism polymermixture envelope of the same or different composition; and apharmaceutical dosage form containing matrix pellets. The matrix pelletreleases acid-soluble drugs by diffusion in acid pH and bydisintegration at pH levels of nominally about 5.0 or higher.

The Aad-GCRA peptides described herein may be formulated in the pHtriggered targeted control release systems described in WO04052339. Theagents described herein may be formulated according to the methodologydescribed in any of WO03105812 (extruded hyrdratable polymers);WO0243767 (enzyme cleavable membrane translocators); WO03007913 andWO3086297 (mucoadhesive systems); WO2072075 (bilayer laminatedformulation comprising pH lowering agent and absorption enhancer);WO04064769 (amidated polypeptides); WO05063156 (solid lipid suspensionwith pseudotropic and/or thixotropic properties upon melting); WO3035029and WO3035041 (erodible, gastric retentive dosage forms); U.S. Pat. No.5,007,790 and U.S. Pat. No. 5,972,389 (sustained release dosage forms);WO041 1271 1 (oral extended release compositions); WO05027878,WO02072033, and WO02072034 (delayed release compositions with natural orsynthetic gum); WO5030182 (controlled release formulations with anascending rate of release); WO05048998 (microencapsulation system); U.S.Pat. No. 5,952,314 (biopolymer); U.S. Pat. No. 5,108,758 (glassy amylosematrix delivery); U.S. Pat. No. 5,840,860 (modified starch baseddelivery). JP10324642 (delivery system comprising chitosan and gastricresistant material such as wheat gliadin or zein); U.S. Pat. No.5,866,619 and U.S. Pat. No. 6,368,629 (saccharide containing polymer);U.S. Pat. No. 6,531,152 (describes a drug delivery system containing awater soluble core (Ca pectinate or other water-insoluble polymers) andouter coat which bursts (e.g. hydrophobic polymer-Eudragrit)); U.S. Pat.No. 6,234,464; U.S. Pat. No. 6,403,130 (coating with polymer containingcasein and high methoxy pectin; WO0174 175 (Maillard reaction product);WO05063206 (solubility increasing formulation); WO040 19872(transferring fusion proteins).

The Aad-GCRA peptides described herein may be formulated usinggastrointestinal retention system technology (GIRES; MerrionPharmaceuticals). GIRES comprises a controlled-release dosage forminside an inflatable pouch, which is placed in a drug capsule for oraladministration. Upon dissolution of the capsule, a gas-generating systeminflates the pouch in the stomach where it is retained for 16-24 hours,all the time releasing agents described herein.

The Aad-GCRA peptides described herein can be formulated in an osmoticdevice including the ones disclosed in U.S. Pat. No. 4,503,030, U.S.Pat. No. 5,609,590 and U.S. Pat. No. 5,358,502. U.S. Pat. No. 4,503,030discloses an osmotic device for dispensing a drug to certain pH regionsof the gastrointestinal tract. More particularly, the invention relatesto an osmotic device comprising a wall formed of a semi-permeable pHsensitive composition that surrounds a compartment containing a drug,with a passageway through the wall connecting the exterior of the devicewith the compartment. The device delivers the drug at a controlled ratein the region of the gastrointestinal tract having a pH of less than3.5, and the device self-destructs and releases all its drug in theregion of the gastrointestinal tract having a pH greater than 3.5,thereby providing total availability for drug absorption. U.S. Pat. Nos.5,609,590 and 5,358,502 disclose an osmotic bursting device fordispensing a beneficial agent to an aqueous environment. The devicecomprises a beneficial agent and osmagent surrounded at least in part bya semi-permeable membrane. The beneficial agent may also function as theosmagent. The semi-permeable membrane is permeable to water andsubstantially impermeable to the beneficial agent and osmagent. Atrigger means is attached to the semi-permeable membrane (e.g., joinstwo capsule halves). The trigger means is activated by a pH of from 3 to9 and triggers the eventual, but sudden, delivery of the beneficialagent. These devices enable the pH-triggered release of the beneficialagent core as a bolus by osmotic bursting.

Exemplary Agents for Combination Therapy

Analgesic Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy with an analgesic agent, e.g., an analgesic compound or ananalgesic polypeptide. These polypeptides and compounds can beadministered with the Aad-GCRA peptides described herein (simultaneouslyor sequentially). They can also be optionally covalently linked orattached to an agent described herein to create therapeutic conjugates.Among the useful analgesic agents are: Calcium channel blockers, 5HTreceptor antagonists (for example 5HT3, 5HT4 and 5HT1 receptorantagonists), opioid receptor agonists (loperamide, fedotozine, andfentanyl), NK1 receptor antagonists, CCK receptor agonists (e.g.,loxiglumide), NK1 receptor antagonists, NK3 receptor antagonists,norepinephrine-serotonin reuptake inhibitors (NSRI), vanilloid andcannabanoid receptor agonists, and sialorphin. Analgesics agents in thevarious classes are described in the literature.

Among the useful analgesic polypeptides are sialorphin-relatedpolypeptides, including those comprising the amino acid sequence QHNPR(SEQ ID NO: 95), including: VQHNPR (SEQ ID NO: 96); VRQHNPR (SEQ ID NO:97); VRGQHNPR (SEQ ID NO: 98); VRGPQHNPR (SEQ ID NO: 99); VRGPRQHNPR(SEQ ID NO: 100); VRGPRRQHNPR (SEQ ID NO: 101); and RQHNPR (SEQ ID NO:102). Sialorphin-related polypeptides bind to neprilysin and inhibitneprilysin-mediated breakdown of substance P and Met-enkephalin. Thus,compounds or polypeptides that are inhibitors of neprilysin are usefulanalgesic agents which can be administered with the polypeptidesdescribed herein in a co-therapy or linked to the polypeptides describedherein, e.g., by a covalent bond. Sialophin and related polypeptides aredescribed in U.S. Pat. No. 6,589,750; U.S. 20030078200 A1; and WO02/051435 A2.

Opioid receptor antagonists and agonists can be administered with theGCRA peptides described herein in co-therapy or linked to the agentdescribed herein, e.g., by a covalent bond. For example, opioid receptorantagonists such as naloxone, naltrexone, methyl nalozone, nalmefene,cypridime, beta funaltrexamine, naloxonazine, naltrindole, andnor-binaltorphimine are thought to be useful in the treatment of IBS. Itcan be useful to formulate opioid antagonists of this type is a delayedand sustained release formulation such that initial release of theantagonist is in the mid to distal small intestine and/or ascendingcolon. Such antagonists are described in WO 01/32180 A2. Enkephalinpentapeptide (HOE825; Tyr-D-Lys-Gly-Phe-L-homoserine) (SEQ ID NO: 103)is an agonist of the mu and delta opioid receptors and is thought to beuseful for increasing intestinal motility {Eur. J. Pharm. 219:445,1992), and this polypeptide can be used in conjunction with thepolypeptides described herein. Also useful is trimebutine which isthought to bind to mu/delta/kappa opioid receptors and activate releaseof motilin and modulate the release of gastrin, vasoactive intestinalpolypeptide, gastrin and glucagons. Kappa opioid receptor agonists suchas fedotozine, asimadoline, and ketocyclazocine, and compounds describedin WO03/097051 and WO05/007626 can be used with or linked to thepolypeptides described herein. In addition, mu opioid receptor agonistssuch as morphine, diphenyloxylate, frakefamide(H-Tyr-D-Ala-Phe(F)-Phe-NH 2 (SEQ ID NO: 104); WO 01/019849 A1) andloperamide can be used.

Tyr-Arg (kyotorphin) is a dipeptide that acts by stimulating the releaseof met-enkephalins to elicit an analgesic effect (J. Biol. Chem262:8165, 1987). Kyotorphin can be used with or linked to the Aad-GCRApeptides described herein.

Chromogranin-derived polypeptide (CgA 47-66; See, e.g., Ghia et al. 2004Regulatory polypeptides 119:199) can be used with or linked to theAad-GCRA peptides described herein.

CCK receptor agonists such as caerulein from amphibians and otherspecies are useful analgesic agents that can be used with or linked tothe Aad-GCRA peptides described herein.

Conotoxin polypeptides represent a large class of analgesic polypeptidesthat act at voltage gated calcium channels, NMDA receptors or nicotinicreceptors. These polypeptides can be used with or linked to thepolypeptides described herein.

Peptide analogs of thymulin (FR Application 2830451) can have analgesicactivity and can be used with or linked to the polypeptides describedherein.

CCK (CCKa or CCKb) receptor antagonists, including loxiglumide anddexloxiglumide (the R-isomer of loxiglumide) (WO 88/05774) can haveanalgesic activity and can be used with or linked to the polypeptidesdescribed herein.

Other useful analgesic agents include 5-HT4 agonists such as tegaserod(Zelnorm®), mosapride, metoclopramide, zacopride, cisapride, renzapride,benzimidazolone derivatives such as BIMU 1 and BIMU 8, and lirexapride.Such agonists are described in: EP1321 142 A1, WO 03/053432A1, EP 505322A1, EP 505322 B1, U.S. Pat. No. 5,510,353, EP 507672 A1, EP 507672 B1,and U.S. Pat. No. 5,273,983.

Calcium channel blockers such as ziconotide and related compoundsdescribed in, for example, EP625162B1, U.S. Pat. No. 5,364,842, U.S.Pat. No. 5,587,454, U.S. Pat. No. 5,824,645, U.S. Pat. No. 5,859,186,U.S. Pat. No. 5,994,305, U.S. Pat. No. 6,087,091, U.S. Pat. No.6,136,786, WO 93/13128 A1, EP 1336409 A1, EP 835126 A1, EP 835126 B1,U.S. Pat. No. 5,795,864, U.S. Pat. No. 5,891,849, U.S. Pat. No.6,054,429, WO 97/01351 A1, can be used with or linked to thepolypeptides described herein.

Various antagonists of the 0, NK-2, and NK-3 receptors (for a review seeGiardina et al. 2003. Drugs 6:758) can be can be used with or linked tothe polypeptides described herein.

NK1 receptor antagonists such as: aprepitant (Merck & Co Inc),vofopitant, ezlopitant (Pfizer, Inc.), R-673 (Hoffmann-La Roche Ltd),SR-48968 (Sanofi Synthelabo), CP-122,721 (Pfizer, Inc.), GW679769 (GlaxoSmith Kline), TAK-637 (Takeda/Abbot), SR-14033, and related compoundsdescribed in, for example, EP 873753 A1, US 20010006972 A1, US20030109417 A1, WO 01/52844 A1, can be used with or linked to thepolypeptides described herein.

NK-2 receptor antagonists such as nepadutant (Menarini Ricerche SpA),saredutant (Sanofi-Synthelabo), GW597599 (Glaxo Smith Kline), SR-144190(Sanoft-Synthelabo) and UK-290795 (Pfizer Inc) can be used with orlinked to the polypeptides described herein.

NK3 receptor antagonists such as osanetant (SR-142801;Sanoft-Synthelabo), SSR-241586, talnetant and related compoundsdescribed in, for example, WO 02/094187 A2, EP 876347 A1, WO 97/21680A1, U.S. Pat. No. 6,277,862, WO 98/1 1090, WO 95/28418, WO 97/19927, andBoden et al. (J Med Chem. 39:1664-75, 1996) can be used with or linkedto the polypeptides described herein.

Norepinephrine-serotonin reuptake inhibitors (NSRI) such as milnacipranand related compounds described in WO 03/077897 A1 can be used with orlinked to the polypeptides described herein.

Vanilloid receptor antagonists such as arvanil and related compoundsdescribed in WO 01/64212 A1 can be used with or linked to thepolypeptides described herein.

The analgesic polypeptides and compounds can be administered with thepolypeptides and agonists described herein (simultaneously orsequentially). The analgesic agents can also be covalently linked to thepolypeptides and agonists described herein to create therapeuticconjugates. Where the analgesic is a polypeptide and is covalentlylinked to an agent described herein the resulting polypeptide may alsoinclude at least one trypsin cleavage site. When present within thepolypeptide, the analgesic polypeptide may be preceded by (if it is atthe carboxy terminus) or followed by (if it is at the amino terminus) atrypsin cleavage site that allows release of the analgesic polypeptide.

In addition to sialorphin-related polypeptides, analgesic polypeptidesinclude: AspPhe, endomorphin-1, endomorphin-2, nocistatin, dalargin,lupron, ziconotide, and substance P.

Agents to Treat Gastrointestinal Disorders

Examples of additional therapeutic agents to treat gastrointestinal andother disorders include agents to treat constipation (e.g., a chloridechannel activator such as the bicylic fatty acid, Lubiprostone (formerlyknown as SPI-0211; Sucampo Pharmaceuticals, Inc.; Bethesda, Md.), alaxative (e.g. a bulk-forming laxative (e.g. nonstarch polysaccharides,Colonel Tablet (polycarbophil calcium), Plantago Ovata®, Equalactin®(Calcium Polycarbophil)), fiber (e.g. FIBERCON® (Calcium Polycarbophil),an osmotic laxative, a stimulant laxative (such as diphenylmethanes(e.g. bisacodyl), anthraquinones (e.g. cascara, senna), and surfactantlaxatives (e.g. castor oil, docusates), an emollient/lubricating agent(such as mineral oil, glycerine, and docusates), MiraLax (BraintreeLaboratories, Braintree Mass.), dexloxiglumide (Forest Laboratories,also known as CR 2017 Rottapharm (Rotta Research Laboratorium SpA)),saline laxatives, enemas, suppositories, and CR 3700 (Rottapharm (RottaResearch Laboratorium SpA); acid reducing agents such as proton pumpinhibitors (e.g., omeprazole (Prilosec®), esomeprazole (Nexium®),lansoprazole (Prevacid®), pantoprazole (Protonix®) and rabeprazole(Aciphex®)) and Histamine H2-receptor antagonist (also known as H2receptor blockers including cimetidine, ranitidine, famotidine andnizatidine); prokinetic agents including itopride, octreotide,bethanechol, metoclopramide (Reglan®), domperidone (Motilium®),erythromycin (and derivatives thereof) or cisapride (Propulsid®);Prokineticin polypeptides homologs, variants and chimeras thereofincluding those described in U.S. Pat. No. 7,052,674 which can be usedwith or linked to the polypeptides described herein; pro-motility agentssuch as the vasostatin-derived polypeptide, chromogranin A (4-16) (See,e.g., Ghia et al. 2004 Regulatory polypeptides 121:31) or motilinagonists (e.g., GM-611 or mitemcinal fumarate) or nociceptin/Orphanin FQreceptor modulators (US20050169917); other peptides which can bind toand/or activate GC-C including those described in US20050287067;complete or partial 5HT (e.g. 5HT1, 5HT2, 5HT3, 5HT4) receptor agonistsor antagonists (including 5HT1A antagonists (e.g. AGI-OO1 (AGItherapeutics), 5HT2B antagonists (e.g. PGN 1091 and PGN1 164 (PharmageneLaboratories Limited), and 5HT4 receptor agonists (such as tegaserod(ZELNORM®), prucalopride, mosapride, metoclopramide, zacopride,cisapride, renzapride, benzimidazolone derivatives such as BIMU 1 andBIMU 8, and lirexapride). Such agonists/modulators are described in:EP1321142 A1, WO 03/053432A1, EP 505322 A1, EP 505322 B1, U.S. Pat. No.5,510,353, EP 507672 A1, EP 507672 B1, U.S. Pat. No. 5,273,983, and U.S.Pat. No. 6,951,867); 5HT3 receptor agonists such as MKC-733; and 5HT3receptor antagonists such as DDP-225 (MCI-225; Dynogen Pharmaceuticals,Inc.), cilansetron (Calmactin®), alosetron (Lotronex®), Ondansetron HCl(Zofran®), Dolasetron (ANZEMET®), palonosetron (Aloxi®), Granisetron(Kytril®), YM060(ramosetron; Astellas Pharma Inc.; ramosetron may begiven as a daily dose of 0.002 to 0.02 mg as described in EP01588707)and ATI-7000 (Aryx Therapeutics, Santa Clara Calif.); muscarinicreceptor agonists; anti-inflammatory agents; antispasmodics includingbut not limited to anticholinergic drugs (like dicyclomine (e.g.Colimex®, Formulex®, Lomine®, Protylol®, Visceral®, Spasmoban®, Bentyl®,Bentylol®), hyoscyamine (e.g. IB-Stat®, Nulev®, Levsin®, Levbid®,Levsinex Timecaps®, Levsin/SL®, Anaspaz®, A-Spas S/L®, Cystospaz®,Cystospaz-M®, Donnamar®, Colidrops Liquid Pediatric®, Gastrosed®, HycoElixir®, Hyosol®, Hyospaz®, Hyosyne®, Losamine®, Medispaz®, Neosol®,Spacol®, Spasdel®, Symax®, Symax SL®), Donnatal (e.g. DonnatalExtentabs®), clidinium (e.g. Quarzan, in combination withLibrium=Librax), methantheline (e.g. Banthine), Mepenzolate (e.g.Cantil), homatropine (e.g. hycodan, Homapin), Propantheline bromide(e.g. Pro-Banthine), Glycopyrrolate (e.g. Robinul®, Robinul Forte®),scopolamine (e.g. Transderm-Scop®, Transderm-V®), hyosine-N-butylbromide(e.g. Buscopan®), Pirenzepine (e.g. Gastrozepin®) Propantheline Bromide(e.g. Propanthel®), dicycloverine (e.g. Merbentyl®), glycopyrroniumbromide (e.g. Glycopyrrolate®), hyoscine hydrobromide, hyoscinemethobromide, methanthelinium, and octatropine); peppermint oil; anddirect smooth muscle relaxants like cimetropium bromide, mebeverine(DUSPATAL®, DUSPATALIN®, COLOFAC MR®, COLOTAL®), otilonium bromide(octilonium), pinaverium (e.g. Dicetel® (pinaverium bromide; Solvay S.A.)), Spasfon® (hydrated phloroglucinol and trimethylphloroglucinol) andtrimebutine (including trimebutine maleate (Modulon®); antidepressants,including but not limited to those listed herein, as well as tricyclicantidepressants like amitriptyline (Elavil®), desipramine (Norpramin®),imipramine (Tofranil®), amoxapine (Asendin®), nortriptyline; theselective serotonin reuptake inhibitors (SSRTs) like paroxetine(Paxil®), fluoxetine (Prozac®), sertraline (Zoloft®), and citralopram(Celexa®); and others like doxepin (Sinequan®) and trazodone (Desyrel®);centrally-acting analgesic agents such as opioid receptor agonists,opioid receptor antagonists (e.g., naltrexone); agents for the treatmentof Inflammatory bowel disease; agents for the treatment of Crohn'sdisease and/or ulcerative colitis (e.g., alequel (Enzo Biochem, Inc.;Farmingsale, N.Y.), the anti-inflammatory polypeptide RDP58 (Genzyme,Inc.; Cambridge, Mass.), and TRAFICET-EN™ (ChemoCentryx, Inc.; SanCarlos, Calif.); agents that treat gastrointestinal or visceral pain;agents that increase cGMP levels (as described in US20040121994) likeadrenergic receptor antagonists, dopamine receptor agonists and PDE(phosphodiesterase) inhibitors including but not limited to thosedisclosed herein; purgatives that draw fluids to the intestine (e.g.,VISICOL®, a combination of sodium phosphate monobasic monohydrate andsodium phosphate dibasic anhydrate); Corticotropin Releasing Factor(CRF) receptor antagonists (including NBI-34041 (Neurocrine Biosciences,San Diego, Calif.), CRH9-41, astressin, R121919 (Janssen Pharmaceutica),CP154,526, NBI-27914, Antalarmin, DMP696 (Bristol-Myers Squibb)CP-316,311 (Pfizer, Inc.), SB723620 (GSK), GW876008 (Neurocrine/GlaxoSmith Kline), ONO-2333Ms (Ono Pharmaceuticals), TS-041 (Janssen), AAG561(Novartis) and those disclosed in U.S. Pat. No. 5,063,245, U.S. Pat. No.5,861,398, US20040224964, US20040198726, US20040176400, US20040171607,US20040110815, US20040006066, and US20050209253); glucagon-likepolypeptides (glp-1) and analogues thereof (including exendin-4 andGTP-010 (Gastrotech Pharma A)) and inhibitors of DPP-IV (DPP-IV mediatesthe inactivation of glp-1); tofisopam, enantiomerically-pureR-tofisopam, and pharmaceutically-acceptable salts thereof (US20040229867); tricyclic anti-depressants of the dibenzothiazepine typeincluding but not limited to Dextoffsopam® (Vela Pharmaceuticals),tianeptine (Stablon®) and other agents described in U.S. Pat. No.6,683,072; (E)-4(1,3bis(cyclohexylmethyl)-1,2,34,-tetrahydro-2,6-diono-9H-purin-8-yl)cinnamicacid nonaethylene glycol methyl ether ester and related compoundsdescribed in WO 02/067942; the probiotic PROBACTRIX® (The BioBalanceCorporation; New York, N.Y.) which contains microorganisms useful in thetreatment of gastrointestinal disorders; antidiarrheal drugs includingbut not limited to loperamide (Imodium, Pepto Diarrhea), diphenoxylatewith atropine (Lomotil, Lomocot), cholestyramine (Questran, Cholybar),atropine (Co-Phenotrope, Diarsed, Diphenoxylate, Lofene, Logen, Lonox,Vi-Atro, atropine sulfate injection) and Xifaxan® (rifaximin; SalixPharmaceuticals Ltd), TZP-201(Tranzyme Pharma Inc.), the neuronalacetylcholine receptor (nAChR) blocker AGI-004 (AGI therapeutics), andbismuth subsalicylate (Pepto-bismol); anxiolytic drugs including but notlimited to Ativan (lorazepam), alprazolam (Xanax®),chlordiazepoxide/clidinium (Librium®, Librax®), clonazepam (Klonopin®),clorazepate (Tranxene®), diazepam (Valium®), estazolam (ProSom®),flurazepam (Dalmane®), oxazepam (Serax®), prazepam (Centrax®), temazepam(Restoril®), triazolam (Halcion®; Bedelix® (Montmorillonite beidellitic;Ipsen Ltd), Solvay SLV332 (ArQuIe Inc), YKP (SK Pharma), Asimadoline(Tioga Pharmaceuticals/Merck), AGI-003 (AGI Therapeutics); neurokininantagonists including those described in US20060040950; potassiumchannel modulators including those described in U.S. Pat. No. 7,002,015;the serotonin modulator AZD7371 (AstraZeneca PIc); M3 muscarinicreceptor antagonists such as darifenacin (Enablex; Novartis AG andzamifenacin (Pfizer); herbal and natural therapies including but notlimited to acidophilus, chamomile tea, evening primrose oil, fennelseeds, wormwood, comfrey, and compounds of Bao-Ji-Wan (magnolol,honokiol, imperatorin, and isoimperatorin) as in U.S. Pat. No.6,923,992; and compositions comprising lysine and an anti-stress agentfor the treatment of irritable bowel syndrome as described in EPO1550443.

Insulin and Insulin Modulating Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy with insulin and related compounds including primate, rodent, orrabbit insulin including biologically active variants thereof includingallelic variants, more preferably human insulin available in recombinantform. Sources of human insulin include pharmaceutically acceptable andsterile formulations such as those available from Eli Lilly(Indianapolis, Ind. 46285) as Humulin™ (human insulin rDNA origin). See,the THE PHYSICIAN'S DESK REFERENCE, 55.sup.th Ed. (2001) MedicalEconomics, Thomson Healthcare (disclosing other suitable humaninsulins).

The Aad-GCRA peptides described herein can also be used in combinationtherapy with agents that can boost insulin effects or levels of asubject upon administration, e.g. glipizide and/or rosiglitazone. Thepolypeptides and agonists described herein can be used in combitherapywith SYMLIN® (pramlintide acetate) and Exenatide® (synthetic exendin-4;a 39 aa polypeptide).

Agents for the Treatment of Postoperative Ileus

The Aad-GCRA peptides described herein can also be used in combinationtherapy with agents (e.g., Entereg™ (alvimopan; formerly called adolor/ADL 8-2698), conivaptan and related agents describe in U.S. Pat. No.6,645,959) used for the treatment of postoperative ileus and otherdisorders.

Anti-Hypertensive Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy with an anti-hypertensive agent including but not limited to:(1) diuretics, such as thiazides, including chlorthalidone,chlorthiazide, dichlorophenamide, hydroflumethiazide, indapamide,polythiazide, and hydrochlorothiazide; loop diuretics, such asbumetanide, ethacrynic acid, furosemide, and torsemide; potassiumsparing agents, such as amiloride, and triamterene; carbonic anhydraseinhibitors, osmotics (such as glycerin) and aldosterone antagonists,such as spironolactone, epirenone, and the like; (2) beta-adrenergicblockers such as acebutolol, atenolol, betaxolol, bevantolol,bisoprolol, bopindolol, carteolol, carvedilol, celiprolol, esmolol,indenolol, metaprolol, nadolol, nebivolol, penbutolol, pindolol,propanolol, sotalol, tertatolol, tilisolol, and timolol, and the like;(3) calcium channel blockers such as amlodipine, aranidipine,azelnidipine, barnidipine, benidipine, bepridil, cinaldipine,clevidipine, diltiazem, efonidipine, felodipine, gallopamil, isradipine,lacidipine, lemildipine, lercanidipine, nicardipine, nifedipine,nilvadipine, nimodepine, nisoldipine, nitrendipine, manidipine,pranidipine, and verapamil, and the like; (4) angiotensin convertingenzyme (ACE) inhibitors such as benazepril; captopril; ceranapril;cilazapril; delapril; enalapril; enalopril; fosinopril; imidapril;lisinopril; losinopril; moexipril; quinapril; quinaprilat; ramipril;perindopril; perindropril; quanipril; spirapril; tenocapril;trandolapril, and zofenopril, and the like; (5) neutral endopeptidaseinhibitors such as omapatrilat, cadoxatril and ecadotril, fosidotril,sampatrilat, AVE7688, ER4030, and the like; (6) endothelin antagonistssuch as tezosentan, A308165, and YM62899, and the like; (7) vasodilatorssuch as hydralazine, clonidine, minoxidil, and nicotinyl alcohol, andthe like; (8) angiotensin II receptor antagonists such as aprosartan,candesartan, eprosartan, irbesartan, losartan, olmesartan, pratosartan,tasosartan, telmisartan, valsartan, and EXP-3137, F16828K, and RNH6270,and the like; (9) α/β adrenergic blockers such as nipradilol, arotinololand amosulalol, and the like; (10) alpha 1 blockers, such as terazosin,urapidil, prazosin, tamsulosin, bunazosin, trimazosin, doxazosin,naftopidil, indoramin, WHP 164, and XENO1O, and the like; (11) alpha 2agonists such as lofexidine, tiamenidine, moxonidine, rilmenidine andguanobenz, and the like; (12) aldosterone inhibitors, and the like; and(13) angiopoietin-2-binding agents such as those disclosed inWO03/030833. Specific anti-hypertensive agents that can be used incombination with polypeptides and agonists described herein include, butare not limited to: diuretics, such as thiazides (e.g., chlorthalidone,cyclothiazide (CAS RN 2259-96-3), chlorothiazide (CAS RN 72956-09-3,which may be prepared as disclosed in U.S. Pat. No. 2,809,194),dichlorophenamide, hydroflumethiazide, indapamide, polythiazide,bendroflumethazide, methyclothazide, polythiazide, trichlormethazide,chlorthalidone, indapamide, metolazone, quinethazone, althiazide (CAS RN5588-16-9, which may be prepared as disclosed in British Patent No.902,658), benzthiazide (CAS RN 91-33-8, which may be prepared asdisclosed in U.S. Pat. No. 3,108,097), buthiazide (which may be preparedas disclosed in British Patent Nos. 861,367), and hydrochlorothiazide),loop diuretics (e.g. bumetanide, ethacrynic acid, furosemide, andtorasemide), potassium sparing agents (e.g. amiloride, and triamterene(CAS Number 396-01-O)), and aldosterone antagonists (e.g. spironolactone(CAS Number 52-01-7), epirenone, and the like); β-adrenergic blockerssuch as Amiodarone (Cordarone, Pacerone), bunolol hydrochloride (CAS RN31969-05-8, Parke-Davis), acebutolol (±N-[3-Acetyl-4-[2-hydroxy-3-[(1methylethyl)amino]propoxy]phenyl]-butanamide, or(±)-3′-Acetyl-4′-[2-hydroxy-3-(isopropylamino) propoxy] butyranilide),acebutolol hydrochloride (e.g. Sectral®, Wyeth-Ayerst), alprenololhydrochloride (CAS RN 13707-88-5 see Netherlands Patent Application No.6,605,692), atenolol (e.g. Tenormin®, AstraZeneca), carteololhydrochloride (e.g. Cartrol® Filmtab®, Abbott), Celiprolol hydrochloride(CAS RN 57470-78-7, also see in U.S. Pat. No. 4,034,009), cetamololhydrochloride (CAS RN 77590-95-5, see also U.S. Pat. No. 4,059,622),labetalol hydrochloride (e.g. Normodyne®, Schering), esmololhydrochloride (e.g. Brevibloc®, Baxter), levobetaxolol hydrochloride(e.g. Betaxon™ Ophthalmic Suspension, Alcon), levobunolol hydrochloride(e.g. Betagan® Liquifilm® with C CAP® Compliance Cap, Allergan), nadolol(e.g. Nadolol, Mylan), practolol (CAS RN 6673-35-4, see also U.S. Pat.No. 3,408,387), propranolol hydrochloride (CAS RN 318-98-9), sotalolhydrochloride (e.g. Betapace AF™, Berlex), timolol (2-Propanol,1-[(1,1-dimethylethyl)amino]-3-[[4-4(4-morpholinyl)-1,2,5-thiadiazol-3-yl]oxy]-,hemihydrate, (S)-, CAS RN 91524-16-2), timolol maleate(S)—I-[(1,1-dimethylethyl)amino]-3-[[4-(4-morpholinyl)-1,2,5-thiadiazol-3-yl] oxy]-2-propanol(Z)-2-butenedioate (1:1) salt, CAS RN 26921-17-5), bisoprolol(2-Propanol,1-[4-[[2-(1-methylethoxy)ethoxy]-methyl]phenoxyl]-3-[(1-meth-ylethyl)amino]-,(±), CAS RN 66722-44-9), bisoprolol fumarate (such as(±)-1-[4-[[2-(1-Methylethoxy)ethoxy]methyl]phenoxy]-3-[(1-methylethyl)amino]-2-propanol(E)-2-butenedioate (2:1) (salt), e.g., Zebeta™, Lederle Consumer),nebivalol (2H-1-Benzopyran-2-methanol,αα′-[iminobis(methylene)]bis[6-fluoro-3,4-dihydro-, CAS RN 99200-09-6see also U.S. Pat. No. 4,654,362), cicloprolol hydrochloride, such2-Propanol,1-[4-[2-(cyclopropylmethoxy)ethoxy]phenoxy]-3-[1-methylethyl)amino]-,hydrochloride, A.A.S. RN 63686-79-3), dexpropranolol hydrochloride(2-Propanol, 1-[1-methylethy)-amino]-3-(1-naphthalenyloxy)-hydrochloride(CAS RN 13071-11-9), diacetolol hydrochloride (Acetamide,N-[3-acetyl-4-[2-hydroxy-3-[(1-methyl-ethyl)amino]propoxy] [phenyl]-,monohydrochloride CAS RN 69796-04-9), dilevalol hydrochloride(Benzamide,2-hydroxy-5-[1-hydroxy-2-[1-methyl-3-phenylpropyl)amino]ethyl]-,monohydrochloride, CAS RN 75659-08-4), exaprolol hydrochloride(2-Propanol, 1-(2-cyclohexylphenoxy)-3-[(1-methylethyl)amino]-,hydrochloride CAS RN 59333-90-3), flestolol sulfate (Benzoic acid,2-fluro-,3-[[2-[aminocarbonyl)amino]-dimethylethyl]amino]-2-hydroxypropylester, (+)-sulfate (1:1) (salt), CAS RN 88844-73-9; metalolhydrochloride (Methanesulfonamide,N-[4-[1-hydroxy-2-(methylamino)propyl]phenyl]-, monohydrochloride CAS RN7701-65-7), metoprolol 2-Propanol,1-[4-(2-methoxyethyl)phenoxy]-3-[1-methylethyl)amino]-; CAS RN37350-58-6), metoprolol tartrate (such as 2-Propanol,1-[4-(2-methoxyethyl)phenoxy]-3-[(1-methylethyl)amino]-, e.g.,Lopressor®, Novartis), pamatolol sulfate (Carbamic acid,[2-[4-[2-hydroxy-3-[(1-methylethyl)amino]propoxyl]phenyl]-ethyl]-,methyl ester, (±) sulfate (salt) (2:1), CAS RN 59954-01-7), penbutololsulfate (2-Propanol,1-(2-cyclopentylphenoxy)-3-[1,1-dimethyle-thyl)amino] 1, (S)—, sulfate(2:1) (salt), CAS RN 38363-32-5), practolol (Acetamide,N-[4-[2-hydroxy-3-[(1-methylethyl)amino]-propoxy]phenyl]-, CAS RN6673-35-4) tiprenolol hydrochloride (Propanol,1-[(1-methylethyl)amino]-3-[2-(methylthio)-phenoxy]-, hydrochloride,(±), CAS RN 39832-43-4), tolamolol (Benzamide,4-[2-[[2-hydroxy-3-(2-methylphenoxy)-propyl] amino] ethoxyl]-, CAS RN38103-61-6), bopindolol, indenolol, pindolol, propanolol, tertatolol,and tilisolol, and the like; calcium channel blockers such as besylatesalt of amlodipine (such as3-ethyl-5-methyl-2-(2-aminoethoxymethyl)-4-(2-chlorophenyl)-,4-dihydro-6-methyl-3,5-pyridinedicarboxylatebenzenesulphonate, e.g., Norvasc®, Pfizer), clentiazem maleate(1,5-Benzothiazepin-4(5H)-one,3-(acetyloxy)-8-chloro-5-[2-(dimethylamino)ethyl]-2,3-dihydro-2-(4-methoxyphenyl)-(2S-cis)-,(Z)-2-butenedioate (1:1), see also U.S. Pat. No. 4,567,195), isradipine(3,5-Pyridinedicarboxylic acid,4-(4-benzofurazanyl)-1,4-dihydro-2,6-dimethyl-, methyl 1-methylethylester,(±)-4(4-benzofurazanyl)-1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate,see also U.S. Pat. No. 4,466,972); nimodipine (such as is isopropyl(2-methoxyethyl) 1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl)-3,5-pyridine-dicarboxylate,e.g. Nimotop®, Bayer), felodipine (such as ethyl methyl4-(2,3-dichlorophenyl)-1,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylate-,e.g. Plendil® Extended-Release, AstraZeneca LP), nilvadipine(3,5-Pyridinedicarboxylic acid,2-cyano-1,4-dihydro-6-methyl-4-(3-nitrophenyl)-,3-methyl5-(1-methylethyl) ester, also see U.S. Pat. No. 3,799,934), nifedipine(such as 3, 5-pyridinedicarboxylicacid,1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, dimethyl ester, e.g.,Procardia XL® Extended Release Tablets, Pfizer), diltiazem hydrochloride(such as 1,5-Benzothiazepin-4(5H)-one,3-(acetyloxy)-5[2-(dimethylamino)ethyl]-2,-3-dihydro-2(4-methoxyphenyl)-,monohydrochloride, (±)-cis., e.g., Tiazac®, Forest), verapamilhydrochloride (such as benzeneacetronitrile,(alpha)-[[3-[[2-(3,4-dimethoxyphenyl)ethyl]methylamino]propyl]-3,4-dimethoxy-(alpha)-(1-methylethyl)hydrochloride, e.g., Isoptin® SR, Knoll Labs), teludipine hydrochloride(3,5-Pyridinedicarboxylic acid,2-[(dimethylamino)methyl]4-[2-[(1E)-3-(1,1-dimethylethoxy)-3-oxo-1-propenyl]phenyl]-1,4-dihydro-6-methyl-,diethyl ester, monohydrochloride) CAS RN 108700-03-4), belfosdil(Phosphonic acid, [2-(2-phenoxy ethyl)-1,3-propane-diyl]bis-, tetrabutylester CAS RN 103486-79-9), fostedil (Phosphonic acid,[[4-(2-benzothiazolyl)phenyl]methyl]-, diethyl ester CAS RN 75889-62-2),aranidipine, azelnidipine, bamidipine, benidipine, bepridil,cinaldipine, clevidipine, efonidipine, gallopamil, lacidipine,lemildipine, lercanidipine, monatepil maleate (1-Piperazinebutanamide,N-(6, 11-dihydrodibenzo(b,e)thiepin-11-yl)₄-(4-fluorophenyl)-, (+)-,(Z)-2-butenedioate (1:1)(±)-N-(6,11-Dihydrodibenzo(b,e)thiep-in-11-yl)-4-(p-fluorophenyl)-1-piperazinebutyramidemaleate (1:1) CAS RN 132046-06-1), nicardipine, nisoldipine,nitrendipine, manidipine, pranidipine, and the like; T-channel calciumantagonists such as mibefradil; angiotensin converting enzyme (ACE)inhibitors such as benazepril, benazepril hydrochloride (such as3-[[1-(ethoxycarbonyl)-3-phenyl-(1S)-propyl]amino]-2,3,4,5-tetrahydro-2-oxo-1H-1-(3S)-benzazepine-1-acetic acid monohydrochloride, e.g., Lotrel®,Novartis), captopril (such as1-[(2S)-3-mercapto-2-methylpropionyl]-L-proline, e.g., Captopril, Mylan,CAS RN 62571-86-2 and others disclosed in U.S. Pat. No. 4,046,889),ceranapril (and others disclosed in U.S. Pat. No. 4,452,790), cetapril(alacepril, Dainippon disclosed in Eur. Therap. Res. 39:671 (1986);40:543 (1986)), cilazapril (Hoffman-LaRoche) disclosed in J. Cardiovasc.Pharmacol. 9:39 (1987), indalapril (delapril hydrochloride(2H-1,2,4-Benzothiadiazine-7-sulfonamide,3-bicyclo[2.2.1]hept-5-en-2-yl-6-chloro-3,4-dihydro-, 1,1-dioxide CAS RN2259-96-3); disclosed in U.S. Pat. No. 4,385,051), enalapril (and othersdisclosed in U.S. Pat. No. 4,374,829), enalopril, enaloprilat,fosinopril, ((such as L-proline,4-cyclohexyl-1-[[[2-methyl-1-(1-oxopropoxy) propoxy](4-phenylbutyl)phosphinyl]acetyl]-, sodium salt, e.g., Monopril, Bristol-Myers Squibband others disclosed in U.S. Pat. No. 4,168,267), fosinopril sodium(L-Proline,4-cyclohexyl-1-[[(R)-[(1S)-2-methyl-1-(1-ox-opropoxy)propox), imidapril,indolapril (Schering, disclosed in J. Cardiovasc. Pharmacol. 5:643, 655(1983)), lisinopril (Merck), losinopril, moexipril, moexiprilhydrochloride (3-Isoquinolinecarboxylic acid,2-[(2S)-2-[[(1S)-1-(ethoxycarbonyl)-3-phenylpropyl]amino]-1-oxopropyl]-1,-2,3,4-tetrahydro-6,7-dimethoxy-,monohydrochloride, (3S)-CAS RN 82586-52-5), quinapril, quinaprilat,ramipril (Hoechsst) disclosed in EP 79022 and Curr. Ther. Res. 40:74(1986), perindopril erbumine (such as2S,3aS,7aS-1-[(S)—N—[(S)-1-Carboxybutyljalanyljhexahydro^-indolinecarboxylicacid, 1-ethyl ester, compound with tert-butylamine (1:1), e.g., Aceon®,Solvay), perindopril (Servier, disclosed in Eur. J. clin. Pharmacol.31:519 (1987)), quanipril (disclosed in U.S. Pat. No. 4,344,949),spirapril (Schering, disclosed in Acta. Pharmacol. Toxicol. 59 (Supp.5): 173 (1986)), tenocapril, trandolapril, zofenopril (and othersdisclosed in U.S. Pat. No. 4,316,906), rentiapril (fentiapril, disclosedin Clin. Exp. Pharmacol. Physiol. 10:131 (1983)), pivopril, YS980,teprotide (Bradykinin potentiator BPP9a CAS RN 35115-60-7), BRL 36,378(Smith Kline Beecham, see EP80822 and EP60668), MC-838 (Chugai, see CA.102:72588v and Jap. J. Pharmacol. 40:373 (1986), CGS 14824 (Ciba-Geigy,3-([1-ethoxycarbonyl-3-phenyl-(1S)-propyl]amino)-2,3,4,5-tetrahydro-2-ox-o-1-(3 S)-benzazepine-1 acetic acid HCl,see U.K. Patent No. 2103614), CGS 16,617 (Ciba-Geigy,3(S)-[[(1S)-5-amino-1-carboxypentyl]amino]-2,3,4,-5-tetrahydro-2-oxo-1H-1-benzazepine-1-ethanoicacid, see U.S. Pat. No. 4,473,575), Ru 44570 (Hoechst, seeArzneimittelforschung 34:1254 (1985)), R 31-2201 (Hoffman-LaRoche seeFEBS Lett. 165:201 (1984)), CI925 (Pharmacologist 26:243, 266 (1984)),WY-44221 (Wyeth, see J. Med. Chem. 26:394 (1983)), and those disclosedin US2003006922 (paragraph 28), U.S. Pat. No. 4,337,201, U.S. Pat. No.4,432,971 (phosphonamidates); neutral endopeptidase inhibitors such asomapatrilat (Vanlev®), CGS 30440, cadoxatril and ecadotril, fasidotril(also known as aladotril or alatriopril), sampatrilat, mixanpril, andgemopatrilat, AVE7688, ER4030, and those disclosed in U.S. Pat. No.5,362,727, U.S. Pat. No. 5,366,973, U.S. Pat. No. 5,225,401, U.S. Pat.No. 4,722,810, U.S. Pat. No. 5,223,516, U.S. Pat. No. 4,749,688, U.S.Pat. No. 5,552,397, U.S. Pat. No. 5,504,080, U.S. Pat. No. 5,612,359,U.S. Pat. No. 5,525,723, EP0599444, EP0481522, EP0599444, EP0595610,EP0534363, EP534396, EP534492, EP0629627; endothelin antagonists such astezosentan, A308165, and YM62899, and the like; vasodilators such ashydralazine (apresoline), clonidine (clonidine hydrochloride(1H-Imidazol-2-amine, N-(2,6-dichlorophenyl)4,5-dihydro-,monohydrochloride CAS RN 4205-91-8), catapres, minoxidil (loniten),nicotinyl alcohol (roniacol), diltiazem hydrochloride (such as1,5-Benzothiazepin-4(5H)-one, 3-(acetyloxy)-5[2-(dimethylamino)ethyl]-2,-3-dihydro-2(4-methoxyphenyl)-,monohydrochloride, (+)-cis, e.g., Tiazac®, Forest), isosorbide dinitrate(such as 1,4:3,6-dianhydro-D-glucitol 2,5-dinitrate e.g., Isordil®Titradose®, Wyeth-Ayerst), sosorbide mononitrate (such as1,4:3,6-dianhydro-D-glucito-1,5-nitrate, an organic nitrate, e.g.,Ismo®, Wyeth-Ayerst), nitroglycerin (such as 2,3 propanetrioltrinitrate, e.g., Nitrostat® Parke-Davis), verapamil hydrochloride (suchas benzeneacetonitrile, (±)-(alpha)[3-[[2-(3,4dimethoxyphenyl)ethyl]methylamino]propyl]-3,4-dimethoxy-(alpha)-(1-methylethyl)hydrochloride, e.g., Covera HS® Extended-Release, Searle), chromonar(which may be prepared as disclosed in U.S. Pat. No. 3,282,938),clonitate (Annalen 1870 155), droprenilamine (which may be prepared asdisclosed in DE2521113), lidoflazine (which may be prepared as disclosedin U.S. Pat. No. 3,267,104); prenylamine (which may be prepared asdisclosed in U.S. Pat. No. 3,152,173), propatyl nitrate (which may beprepared as disclosed in French Patent No. 1,103,113), mioflazinehydrochloride (1-Piperazineacetamide,3-(aminocarbonyl)₄-[4,4-bis(4-fluorophenyl)butyl]-N-(2,6-dichlorophenyl)-,dihydrochloride CAS RN 83898-67-3), mixidine (Benzeneethanamine,3,4-dimethoxy-N-(1-methyl-2-pyrrolidinylidene)-Pyrrolidine,2-[(3,4-dimethoxyphenethyl)imino]-1-methyl-1-Methyl-2-[(3,4-dimethoxyphenethyl)imino]pyrrolidine CAS RN 27737-38-8), molsidomine(1,2,3-Oxadiazolium, 5-[(ethoxycarbonyl)amino]-3-(4-morpholinyl)-, innersalt CAS RN 25717-80-0), isosorbide mononitrate (D-Glucitol,1,4:3,6-dianhydro-, 5-nitrate CAS RN 16051-77-7), erythrityltetranitrate (1,2,3,4-Butanetetrol, tetranitrate, (2R,3S)-rel-CAS RN7297-25-8), clonitrate (1,2-Propanediol, 3-chloro-, dinitrate (7CI, 8CI,9CI) CAS RN 2612-33-1), dipyridamole Ethanol,2,2′,2″,2″″-[(4,8-di-1-piperidinylpyrimido[5,4-d]pyrimidine-2,6-diyl)dinitrilo]tetrakis-CASRN 58-32-2), nicorandil (CAS RN 65141-46-0 3-), pyridinecarboxamide(N-[2-(nitrooxy)ethyl]-Nisoldipine3,5-Pyridinedicarboxylic acid,1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, methyl 2-methylpropyl esterCAS RN 63675-72-9), nifedipine3,5-Pyridinedicarboxylic acid,1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-, dimethyl ester CAS RN21829-25-4), perhexiline maleate (Piperidine,2-(2,2-dicyclohexylethyl)-, (2Z)-2-butenedioate (1:1) CAS RN 6724-53-4),oxprenolol hydrochloride (2-Propanol,1-[(1-methylethyl)amino]-3-[2-(2-propenyloxy)phenoxy]-, hydrochlorideCAS RN 6452-73-9), pentrinitrol (1,3-Propanediol,2,2-bis[(nitrooxy)methyl]-, mononitrate (ester) CAS RN 1607-17-6),verapamil (Benzeneacetonitrile,α-[3-[[2-(3,4-dimethoxyphenyl)ethyl]-methylamino]propyl]-3,4-dimethoxy-α-(1-methylethyl)-CAS RN 52-53-9) and the like; angiotensinII receptor antagonists such as, aprosartan, zolasartan, olmesartan,pratosartan, F16828K, RNH6270, candesartan(1H-Benzimidazole-7-carboxylic acid,2-ethoxy-1-[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]4-yl]methyl]-CAS RN139481-59-7), candesartan cilexetil((+/−)-1-(cyclohexylcarbonyloxy)ethyl-2-ethoxy-1-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]-1H-benzimidazolecarboxylate, CAS RN 145040-37-5, U.S. Pat. No. 5,703,110 and U.S. Pat.No. 5,196,444), eprosartan(3-[1-4-carboxyphenylmethyl)-2-n-butyl-imidazol-5-yl]-(2-thienylmethyl)propenoic acid, U.S. Pat. No. 5,185,351 and U.S. Pat. No. 5,650,650),irbesartan (2-n-butyl-3-[[2′-(1h-tetrazol-5-yl)biphenyl-4-yl]methyl]1,3-diazazspiro[4,4]non-1-en-4-one, U.S. Pat. No. 5,270,317 and U.S.Pat. No. 5,352,788), losartan(2-N-butyl-4-chloro-5-hydroxymethyl-1-[(2′-(1H-tetrazol-5-yl)biphenyl-4-yl)-methyl]imidazole,potassium salt, U.S. Pat. No. 5,138,069, U.S. Pat. No. 5,153,197 andU.S. Pat. No. 5,128,355), tasosartan(5,8-dihydro-2,4-dimethyl-8-[(2′-(1H-tetrazol-5-yl)[1,r-biphenyl]4-yl)methyl]-pyrido[2,3-d]pyrimidin-7(6H)-one,U.S. Pat. No. 5,149,699), telmisartan(4′-[(1,4-dimethyl-2′-propyl-(2,6′-bi-1H-benzimidazol)-r-yl)]-[1,1′-biphenyl]-2-carboxylic acid, CAS RN 144701-48-4, U.S. Pat. No.5,591,762), milfasartan, abitesartan, valsartan (Diovan® (Novartis),(S)—N-valeryl-N-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl)methyl]valine, U.S.Pat. No. 5,399,578), EXP-3137(2-N-butyl-4-chloro-1-[(2′-(1H-tetrazol-5-yl)biphenyl-4-yl)-methyl]imidazole-5-carboxylicacid, U.S. Pat. No. 5,138,069, U.S. Pat. No. 5,153,197 and U.S. Pat. No.5,128,355),3-(2′-(tetrazol-5-yl)-1,r-biphen-4-yl)methyl-5,7-dimethyl-2-ethyl-3H-imidazo[4,5-b]pyridine,4′[2-ethyl-4-methyl-6-(5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-2-yl]-benzimidazol-1-yl]-methyl]-1,r-biphenyl]-2-carboxylicacid,2-butyl-6-(1-methoxy-1-methylethyl)-2-[2′-)IH-tetrazol-5-yl)biphenyl-4-ylmethyl]guinazolin-4(3H)-one,3-[2′-carboxybiphenyl-4-yl)methyl]-2-cyclopropyl-7-methyl-3H-imidazo[4,5-b]pyridine,2-butyl-4-chloro-1-[(2′-tetrazol-5-yl)biphenyl-4-yl)methyl]imidazole-carboxylicacid, 2-butyl-4-chloro-1-[[2′-(1H-tetrazol-5-yl) [1,1′-biphenyl]-4-yl]methyl]-1H-imidazole-5-carboxylicacid-1-(ethoxycarbonyl-oxy)ethyl ester potassium salt, dipotassium2-butyl-4-(methylthio)-1-[[2-[[[(propylamino)carbonyl]amino]-sulfonyl](1,1′-biphenyl)-4-yl]methyl]-1H-imidazole-5-carboxylate,methyl-2-[[4-butyl-2-methyl-6-oxo-5-[[2′-(1H-tetrazol-5-yl)-[1,1′-biphenyl]-4-yl]methyl]-1-(6H)-pyrimidinyl]methyl]-3-thiophencarboxylate,5-[(3,5-dibutyl-1H-1,2,4-triazol-1-yl)methyl]-2-[2-(1H-tetrazol-5-ylphenyl)]pyridine,6-butyl-2-(2-phenylethyl)-5 [[2′-(IH-tetrazol-5-yl)[1,1′-biphenyl]-4-methyl]pyrimidin-4-(3H)-one D,L lysinesalt,5-methyl-7-n-propyl-8-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-[1,2,4]-triazolo[1,5-c]pyrimidin-2(3H)-one,2,7-diethyl-5-[[2′-(5-tetrazoly)biphenyl-4-yl]methyl]-5H-pyrazolo[1,5-b][1,2,4]triazolepotassium salt,2-[2-butyl-4,5-dihydro-4-oxo-3-[2′-(1H-tetrazol-5-yl)-4-biphenylmethyl]-3H-imidazol[4,5-c]pyridine-5-ylmethyl]benzoicacid, ethyl ester, potassium salt,3-methoxy-2,6-dimethyl-4-[[2′(1H-tetrazol-5-yl)-1,1′-biphenyl-4-yl]methoxy]pyridine,2-ethoxy-1-[[2′-(5-oxo-2,5-dihydro-1,2,4-oxadiazol-3-yl)biphenyl-4-yl]methyl]-1H-benzimidazole-7-carboxylicacid,1-[N-(2′-(1H-tetrazol-5-yl)biphenyl-4-yl-methyl)-N-valerolylaminomethyl)cyclopentane-1-carboxylicacid, 7-methyl-2n-propyl-3-[[2′1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-3H-imidazo[4,5-6]pyridine,2-[5-[(2-ethyl-5,7-dimethyl-3H-imidazo[4,5-b]pyridine-3-yl)methyl]-2-quinolinyl]sodiumbenzoate, 2-butyl-6-chloro-4-hydroxymethyl-5-methyl-3-[[2′-(IH-tetrazol-5-yl)biphenyl-4-yl]methyl]pyridine,2-[[[2-butyl-1-[(4-carboxyphenyl)methyl]-1H-imidazol-5-yl]methyl]amino]benzoicacid tetrazol-5-yl)biphenyl-4-yl]methyl]pyrimidin-6-one,4(S)-[4-(carboxymethyl)phenoxy]-N-[2(R)-[4-(2-sulfobenzamido)imidazol-1-yl]octanoyl]-L-proline,1-(2,6-dimethylphenyl)-4-butyl-1,3-dihydro-3-[[6-[2-(1H-tetrazol-5-yl)phenyl]-3-pyridinyl]methyl]-2H-imidazol-2-one,5,8-ethano-5,8-dimethyl-2-n-propyl-5,6,7,8-tetrahydro-1-[[2′(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-1H,4H-1,3,4a,8a-tetrazacyclopentanaphthalene-9-one,4-[1-[2′-(1,2,3,4-tetrazol-5-yl)biphen-4-yl)methylamino]-5,6,7,8-tetrahydro-2-trifylquinazoline,2-(2-chlorobenzoyl)imino-5-ethyl-3-[2′-(1H-tetrazole-5-yl)biphenyl-4-yl)methyl-1,3,4-thiadiazoline,2-[5-ethyl-3-[2-(1H-tetrazole-5-yl)biphenyl-4-yl]methyl-1,3,4-thiazoline-2-ylidene]aminocarbonyl-1-cyclopentencarboxylicacid dipotassium salt, and2-butyl-4-[N-methyl-N-(3-methylcrotonoyl)amino]-1-[[2′-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl]-1H-imidzole-5-carboxylicacid 1-ethoxycarbonyloxyethyl ester, those disclosed in patentpublications EP475206, EP497150, EP539086, EP539713, EP535463, EP535465,EP542059, EP497121, EP535420, EP407342, EP415886, EP424317, EP435827,EP433983, EP475898, EP490820, EP528762, EP324377, EP323841, EP420237,EP500297, EP426021, EP480204, EP429257, EP430709, EP434249, EP446062,EP505954, EP524217, EP514197, EP514198, EP514193, EP514192, EP450566,EP468372, EP485929, EP503162, EP533058, EP467207 EP399731, EP399732,EP412848, EP453210, EP456442, EP470794, EP470795, EP495626, EP495627,EP499414, EP499416, EP499415, EP511791, EP516392, EP520723, EP520724,EP539066, EP438869, EP505893, EP530702, EP400835, EP400974, EP401030,EP407102, EP411766, EP409332, EP412594, EP419048, EP480659, EP481614,EP490587, EP467715, EP479479, EP502725, EP503838, EP505098, EP505111EP513,979 EP507594, EP510812, EP511767, EP512675, EP512676, EP512870,EP517357, EP537937, EP534706, EP527534, EP540356, EP461040, EP540039,EP465368, EP498723, EP498722, EP498721, EP515265, EP503785, EP501892,EP519831, EP532410, EP498361, EP432737, EP504888, EP508393, EP508445,EP403159, EP403158, EP425211, EP427463, EP437103, EP481448, EP488532,EP501269, EP500409, EP540400, EP005528, EP028834, EP028833, EP411507,EP425921, EP430300, EP434038, EP442473, EP443568, EP445811, EP459136,EP483683, EP518033, EP520423, EP531876, EP531874, EP392317, EP468470,EP470543, EP502314, EP529253, EP543263, EP540209, EP449699, EP465323,EP521768, EP415594, WO92/14468, WO93/08171, WO93/08169, WO91/00277,WO91/00281, WO91/14367, WO92/00067, WO92/00977, WO92/20342, WO93/04045,WO93/04046, WO91/15206, WO92/14714, WO92/09600, WO92/16552, WO93/05025,WO93/03018, WO91/07404, WO92/02508, WO92/13853, WO91/19697, WO91/11909,WO91/12001, WO91/11999, WO91/15209, WO91/15479, WO92/20687, WO92/20662,WO92/20661, WO93/01177, WO91/14679, WO91/13063, WO92/13564, WO91/17148,WO91/18888, WO91/19715, WO92/02257, WO92/04335, WO92/05161, WO92/07852,WO92/15577, WO93/03033, WO91/16313, WO92/00068, WO92/02510, WO92/09278,WO9210179, WO92/10180, WO92/10186, WO92/10181, WO92/10097, WO92/10183,WO92/10182, WO92/10187, WO92/10184, WO92/10188, WO92/10180, WO92/10185,WO92/20651, WO93/03722, WO93/06828, WO93/03040, WO92/19211, WO92/22533,WO92/06081, WO92/05784, WO93/00341, WO92/04343, WO92/04059, U.S. Pat.No. 5,104,877, U.S. Pat. No. 5,187,168, U.S. Pat. No. 5,149,699, U.S.Pat. No. 5,185,340, U.S. Pat. No. 4,880,804, U.S. Pat. No. 5,138,069,U.S. Pat. No. 4,916,129, U.S. Pat. No. 5,153,197, U.S. Pat. No.5,173,494, U.S. Pat. No. 5,137,906, U.S. Pat. No. 5,155,126, U.S. Pat.No. 5,140,037, U.S. Pat. No. 5,137,902, U.S. Pat. No. 5,157,026, U.S.Pat. No. 5,053,329, U.S. Pat. No. 5,132,216, U.S. Pat. No. 5,057,522,U.S. Pat. No. 5,066,586, U.S. Pat. No. 5,089,626, U.S. Pat. No.5,049,565, U.S. Pat. No. 5,087,702, U.S. Pat. No. 5,124,335, U.S. Pat.No. 5,102,880, U.S. Pat. No. 5,128,327, U.S. Pat. No. 5,151,435, U.S.Pat. No. 5,202,322, U.S. Pat. No. 5,187,159, U.S. Pat. No. 5,198,438,U.S. Pat. No. 5,182,288, U.S. Pat. No. 5,036,048, U.S. Pat. No.5,140,036, U.S. Pat. No. 5,087,634, U.S. Pat. No. 5,196,537, U.S. Pat.No. 5,153,347, U.S. Pat. No. 5,191,086, U.S. Pat. No. 5,190,942, U.S.Pat. No. 5,177,097, U.S. Pat. No. 5,212,177, U.S. Pat. No. 5,208,234,U.S. Pat. No. 5,208,235, U.S. Pat. No. 5,212,195, U.S. Pat. No.5,130,439, U.S. Pat. No. 5,045,540, U.S. Pat. No. 5,041,152, and U.S.Pat. No. 5,210,204, and pharmaceutically acceptable salts and estersthereof; α/β adrenergic blockers such as nipradilol, arotinolol,amosulalol, bretylium tosylate (CAS RN: 61-75-6), dihydroergtaminemesylate (such asergotaman-3′,6′,18-trione,9,-10-dihydro-12′-hydroxy-2′-methyl-5′-(phenylmethyl)-(5′(α))-,monomethanesulfonate, e.g., DHE 45® Injection, Novartis), carvedilol(such as (±)-1-(Carbazol-4-yloxy)-3-[[2-(o-methoxyphenoxy)ethyl]amino]-2-propanol, e.g., Coreg®, SmithKline Beecham), labetalol (such as5-[1-hydroxy-2-[(1-methyl-3-phenylpropyl) amino]ethyljsalicylamidemonohydrochloride, e.g., Normodyne®, Schering), bretylium tosylate(Benzenemethanaminium, 2-bromo-N-ethyl-N,N-dimethyl-, salt with4-methylbenzenesulfonic acid (1:1) CAS RN 61-75-6), phentolaminemesylate (Phenol,3-[[(4,5-dihydro-1H-imidazol-2-yl)methyl](4-methylphenyl)amino]-,monomethanesulfonate (salt) CAS RN 65-28-1), solypertine tartrate(5H-1,3-Dioxolo[4,5-f]indole,7-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-,(2R,3R)-2,3-dihydroxybutanedioate (1:1) CAS RN 5591-43-5), zolertinehydrochloride (Piperazine, 1-phenyl4-[2-(1H-tetrazol-5-yl)ethyl]-,monohydrochloride (8Cl, 9Cl) CAS RN 7241-94-3) and the like; aadrenergic receptor blockers, such as alfuzosin (CAS RN: 81403-68-1),terazosin, urapidil, prazosin (Minipress®), tamsulosin, bunazosin,trimazosin, doxazosin, naftopidil, indoramin, WHP 164, XENO1O,fenspiride hydrochloride (which may be prepared as disclosed in U.S.Pat. No. 3,399,192), proroxan (CAS RN 33743-96-3), and labetalolhydrochloride and combinations thereof; α 2 agonists such as methyldopa,methyldopa HCL, lofexidine, tiamenidine, moxonidine, rilmenidine,guanobenz, and the like; aldosterone inhibitors, and the like; renininhibitors including Aliskiren (SPP1OO; Novartis/Speedel);angiopoietin-2-binding agents such as those disclosed in WO03/030833;anti-angina agents such as ranolazine (hydrochloride1-Piperazineacetamide,N-(2,6-dimethylphenyl)-4-[2-hydroxy-3-(2-methoxyphenoxy)propyl]-,dihydrochloride CAS RN 95635-56-6), betaxolol hydrochloride (2-Propanol,1-[4-[2 (cyclopropylmethoxy)ethyl]phenoxy]-3-[(1-methylethyl)amino]-,hydrochloride CAS RN 63659-19-8), butoprozine hydrochloride (Methanone,[4-[3(dibutylamino)propoxy]phenyl](2-ethyl-3-indolizinyl)-,monohydrochloride CAS RN 62134-34-3), cinepazetmaleatel-Piperazineacetic acid,4-[1-oxo-3-(3,4,5-trimethoxyphenyl)-2-propenyl]-, ethyl ester,(2Z)-2-butenedioate (1:1) CAS RN 50679-07-7), tosifen(Benzenesulfonamide, 4-methyl-N-[[[(1S)-1-methyl-2-phenylethyl]amino]carbonyl]-CAS RN 32295-184), verapamilhydrochloride(Benzeneacetonitrile,α-[3-[[2-(3,4-dimethoxyphenyl)ethyl]methylamino]propyl]-3,4-dimethoxy-α-(1-methylethyl)-,monohydrochloride CAS RN 152-114), molsidomine (1,2,3-Oxadiazolium,5-[(ethoxycarbonyl)amino]-3-(4-morpholinyl)-, inner salt CAS RN25717-80-0), and ranolazine hydrochloride (1-Piperazineacetamide,N-(2,6-dimethylphenyl)₄-[2-hydroxy-3-(2-meth-oxyphenoxy)propyl]-,dihydrochloride CAS RN 95635-56-6); tosifen (Benzenesulfonamide,4-methyl-N-[[[(1S)-1-methyl-2-phenylethyl]amino] carbonyl]-CAS RN32295-184); adrenergic stimulants such as guanfacine hydrochloride (suchas N-amidino-2-(2,6-dichlorophenyl) acetamide hydrochloride, e.g.,Tenex® Tablets available from Robins); methyldopa-hydrochlorothiazide(such as levo-3-(3,4-dihydroxyphenyl)-2-methylalanine) combined withHydrochlorothiazide (such as6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide1,1-dioxide, e.g., the combination as, e.g., Aldoril® Tablets availablefrom Merck), methyldopa-chlorothiazide (such as 6-chloro-2H-1,2,4-benzothiadiazine-7-sulfonamide 1,1-dioxide and methyldopa asdescribed above, e.g., Aldoclor®, Merck), clonidine hydrochloride (suchas 2-(2,6-dichlorophenylamino)-2-imidazoline hydrochloride andchlorthalidone (such as 2-chloro-5-(1-hydroxy-3-oxo-1-isoindolinyl)benzenesulfonamide), e.g., Combipres®, Boehringer Ingelheim), clonidinehydrochloride (such as 2-(2,6-dichlorophenylamino)-2-imidazolinehydrochloride, e.g., Catapres®, Boehringer Ingelheim), clonidine(1H-Imidazol-2-amine, N-(2,6-dichlorophenyl)4,5-dihydro-CAS RN4205-90-7), Hyzaar (Merck; a combination of losartan andhydrochlorothiazide), Co-Diovan (Novartis; a combination of valsartanand hydrochlorothiazide, Lotrel (Novartis; a combination of benazepriland amlodipine) and Caduet (Pfizer; a combination of amlodipine andatorvastatin), and those agents disclosed in US20030069221.

Agents for the Treatment of Respiratory Disorders

The Aad-GCRA peptides described herein can be used in combinationtherapy with one or more of the following agents useful in the treatmentof respiratory and other disorders including but not limited to: (1)β-agonists including but not limited to: albuterol (PRO VENTIL®, S ALBUTAMOl®, VENTOLIN®), bambuterol, bitoterol, clenbuterol, fenoterol,formoterol, isoetharine (BRONKOSOL®, BRONKOMETER®), metaproterenol(ALUPENT®, METAPREL®), pirbuterol (MAXAIR®), reproterol, rimiterol,salmeterol, terbutaline (BRETHAIRE®, BRETHINE®, BRICANYL®), adrenalin,isoproterenol (ISUPREL®), epinephrine bitartrate (PRIMATENE®),ephedrine, orciprenline, fenoterol and isoetharine; (2) steroids,including but not limited to beclomethasone, beclomethasonedipropionate, betamethasone, budesonide, bunedoside, butixocort,dexamethasone, flunisolide, fluocortin, fluticasone, hydrocortisone,methyl prednisone, mometasone, predonisolone, predonisone, tipredane,tixocortal, triamcinolone, and triamcinolone acetonide; (3)β2-agonist-corticosteroid combinations [e.g., salmeterol-fluticasone (ADV AIR®), formoterol-budesonid (S YMBICORT®)]; (4) leukotriene D4receptor antagonists/leukotriene antagonists/LTD4 antagonists (i.e., anycompound that is capable of blocking, inhibiting, reducing or otherwiseinterrupting the interaction between leukotrienes and the Cys LTIreceptor) including but not limited to: zafhiukast, montelukast,montelukast sodium (SINGULAIR®), pranlukast, iralukast, pobilukast,SKB-106,203 and compounds described as having LTD4 antagonizing activitydescribed in U.S. Pat. No. 5,565,473; (5) 5-lipoxygenase inhibitorsand/or leukotriene biosynthesis inhibitors [e.g., zileuton and BAY1005(CA registry 128253-31-6)]; (6) histamine H1 receptorantagonists/antihistamines (i.e., any compound that is capable ofblocking, inhibiting, reducing or otherwise interrupting the interactionbetween histamine and its receptor) including but not limited to:astemizole, acrivastine, antazoline, azatadine, azelastine, astamizole,bromopheniramine, bromopheniramine maleate, carbinoxamine, carebastine,cetirizine, chlorpheniramine, chloropheniramine maleate, cimetidineclemastine, cyclizine, cyproheptadine, descarboethoxyloratadine,dexchlorpheniramine, dimethindene, diphenhydramine, diphenylpyraline,doxylamine succinate, doxylamine, ebastine, efletirizine, epinastine,famotidine, fexofenadine, hydroxyzine, hydroxyzine, ketotifen,levocabastine, levocetirizine, levocetirizine, loratadine, meclizine,mepyramine, mequitazine, methdilazine, mianserin, mizolastine,noberastine, norasternizole, noraztemizole, phenindamine, pheniramine,picumast, promethazine, pynlamine, pyrilamine, ranitidine, temelastine,terfenadine, trimeprazine, tripelenamine, and triprolidine; (7) ananticholinergic including but not limited to: atropine, benztropine,biperiden, flutropium, hyoscyamine (e.g. Levsin®; Levbid®; Levsin/SL®,Anaspaz®, Levsinex Timecaps®, NuLev®), ilutropium, ipratropium,ipratropium bromide, methscopolamine, oxybutinin, rispenzepine,scopolamine, and tiotropium; (8) an anti-tussive including but notlimited to: dextromethorphan, codeine, and hydromorphone; (9) adecongestant including but not limited to: pseudoephedrine andphenylpropanolamine; (10) an expectorant including but not limited to:guafenesin, guaicolsulfate, terpin, ammonium chloride, glycerolguaicolate, and iodinated glycerol; (11) a bronchodilator including butnot limited to: theophylline and aminophylline; (12) ananti-inflammatory including but not limited to: fluribiprofen,diclophenac, indomethacin, ketoprofen, S-ketroprophen, tenoxicam; (13) aPDE (phosphodiesterase) inhibitor including but not limited to thosedisclosed herein; (14) a recombinant humanized monoclonal antibody [e.g.xolair (also called omalizumab), rhuMab, and talizumab]; (15) ahumanized lung surfactant including recombinant forms of surfactantproteins SP-B, SP-C or SP-D [e.g. SURFAXIN®, formerly known as dsc-104(Discovery Laboratories)], (16) agents that inhibit epithelial sodiumchannels (ENaC) such as amiloride and related compounds; (17)antimicrobial agents used to treat pulmonary infections such asacyclovir, amikacin, amoxicillin, doxycycline, trimethoprinsulfamethoxazole, amphotericin B, azithromycin, clarithromycin,roxithromycin, clarithromycin, cephalosporins (ceffoxitin, cefmetazoleetc), ciprofloxacin, ethambutol, gentimycin, ganciclovir, imipenem,isoniazid, itraconazole, penicillin, ribavirin, rifampin, rifabutin,amantadine, rimantidine, streptomycin, tobramycin, and vancomycin; (18)agents that activate chloride secretion through Ca++ dependent chloridechannels (such as purinergic receptor (P2Y(2) agonists); (19) agentsthat decrease sputum viscosity, such as human recombinant DNase 1,(Pulmozyme®); (20) nonsteroidal anti-inflammatory agents (acemetacin,acetaminophen, acetyl salicylic acid, alclofenac, alminoprofen, apazone,aspirin, benoxaprofen, bezpiperylon, bucloxic acid, carprofen, clidanac,diclofenac, diclofenac, diflunisal, diflusinal, etodolac, fenbufen,fenbufen, fenclofenac, fenclozic acid, fenoprofen, fentiazac, feprazone,flufenamic acid, flufenisal, flufenisal, fluprofen, flurbiprofen,flurbiprofen, furofenac, ibufenac, ibuprofen, indomethacin,indomethacin, indoprofen, isoxepac, isoxicam, ketoprofen, ketoprofen,ketorolac, meclofenamic acid, meclofenamic acid, mefenamic acid,mefenamic acid, miroprofen, mofebutazone, nabumetone oxaprozin,naproxen, naproxen, niflumic acid, oxaprozin, oxpinac, oxyphenbutazone,phenacetin, phenylbutazone, phenylbutazone, piroxicam, piroxicam,pirprofen, pranoprofen, sudoxicam, tenoxican, sulfasalazine, sulindac,sulindac, suprofen, tiaprofenic acid, tiopinac, tioxaprofen, tolfenamicacid, tolmetin, tolmetin, zidometacin, zomepirac, and zomepirac); and(21) aerosolized antioxidant therapeutics such as S-Nitrosoglutathione.

Anti-Obesity Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy with an anti-obesity agent. Suitable such agents include, butare not limited to: 1 1β HSD-I (11-beta hydroxy steroid dehydrogenasetype 1) inhibitors, such as BVT 3498, BVT 2733,3-(1-adamantyl)-4-ethyl-5-(ethylthio)-4H-1,2,4-triazole,3-(1-adamantyl)-5-(3,4,5-trimethoxyphenyl)-4-methyl-4H-1,2,4-triazole,3-adamantanyl-4,5,6,7,8,9,10,11,12,3a-decahydro-1,2,4-triazolo[4,3-a][11]annulene, and those compounds disclosed in WO01/90091, WO01/90090,WO01/90092 and WO02/072084; 5HT antagonists such as those inWO03/037871, WO03/037887, and the like; 5HTIa modulators such ascarbidopa, benserazide and those disclosed in U.S. Pat. No. 6,207,699,WO03/031439, and the like; 5HT2c (serotonin receptor 2c) agonists, suchas BVT933, DPCA37215, IK264, PNU 22394, WAY161503, R-1065, SB 243213(Glaxo Smith Kline) and YM 348 and those disclosed in U.S. Pat. No.3,914,250, WO00/77010, WO02/36596, WO02/48124, WO02/10169, WO01/66548,WO02/44152, WO02/51844, WO02/40456, and WO02/40457; 5HT6 receptormodulators, such as those in WO03/030901, WO03/035061, WO03/039547, andthe like; acyl-estrogens, such as oleoyl-estrone, disclosed in delMar-Grasa, M. et al, Obesity Research, 9:202-9 (2001) and JapanesePatent Application No. JP 2000256190; anorectic bicyclic compounds suchas 1426 (Aventis) and 1954 (Aventis), and the compounds disclosed inWO00/18749, WO01/32638, WO01/62746, WO01/62747, and WO03/015769; CB 1(cannabinoid-1 receptor) antagonist/inverse agonists such as rimonabant(Acomplia; Sanofi), SR-147778 (Sanofi), SR-141716 (Sanofi), BAY 65-2520(Bayer), and SLV 319 (Solvay), and those disclosed in patentpublications U.S. Pat. No. 4,973,587, U.S. Pat. No. 5,013,837, U.S. Pat.No. 5,081,122, U.S. Pat. No. 5,112,820, U.S. Pat. No. 5,292,736, U.S.Pat. No. 5,532,237, U.S. Pat. No. 5,624,941, U.S. Pat. No. 6,028,084,U.S. Pat. No. 6,509,367, U.S. Pat. No. 6,509,367, WO96/33159,WO97/29079, WO98/31227, WO98/33765, WO98/37061, WO98/41519, WO98/43635,WO98/43636, WO99/02499, WO00/10967, WO00/10968, WO01/09120, WO01/58869,WO01/64632, WO01/64633, WO01/64634, WO01/70700, WO01/96330, WO02/076949,WO03/006007, WO03/007887, WO03/020217, WO03/026647, WO03/026648,WO03/027069, WO03/027076, WO03/027114, WO03/037332, WO03/040107,WO03/086940, WO03/084943 and EP658546; CCK-A (cholecystokinin-A)agonists, such as AR-R 15849, GI 181771 (GSK), JMV-180, A-71378, A-71623and SR146131 (Sanofi), and those described in U.S. Pat. No. 5,739,106;CNTF (Ciliary neurotrophic factors), such as GI-181771(Glaxo-SmithKline), SRI 46131 (Sanofi Synthelabo), butabindide, PD170,292, and PD 149164 (Pfizer); CNTF derivatives, such as Axokine®(Regeneron), and those disclosed in WO94/09134, WO98/22128, andWO99/43813; dipeptidyl peptidase IV (DP-IV) inhibitors, such asisoleucine thiazolidide, valine pyrrolidide, NVP-DPP728, LAF237, P93/01,P 3298, TSL 225 (tryptophyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylicacid; disclosed by Yamada et al, Bioorg. & Med. Chem. Lett. 8 (1998)1537-1540), TMC-2A/2B/2C, CD26 inhibtors, FE 999011, P9310/K364, VIP0177, SDZ 274-444, 2-cyanopyrrolidides and 4-cyanopyrrolidides asdisclosed by Ashworth et al, Bioorg. & Med. Chem. Lett., Vol. 6, No. 22,pp 1163-1166 and 2745-2748 (1996) and the compounds disclosed patentpublications. WO99/38501, WO99/46272, WO99/67279 (Probiodrug),WO99/67278 (Probiodrug), WO99/61431 (Probiodrug), WO02/083128,WO02/062764, WO03/000180, WO03/000181, WO03/000250, WO03/002530,WO03/002531, WO03/002553, WO03/002593, WO03/004498, WO03/004496,WO03/017936, WO03/024942, WO03/024965, WO03/033524, WO03/037327 andEP1258476; growth hormone secretagogue receptor agonists/antagonists,such as NN703, hexarelin, MK-0677 (Merck), SM-130686, CP-424391(Pfizer), LY 444,711 (Eli Lilly), L-692,429 and L-163,255, and such asthose disclosed in U.S. Ser. No. 09/662,448, U.S. provisionalapplication 60/203,335, U.S. Pat. No. 6,358,951, US2002049196,US2002/022637, WO01/56592 and WO02/32888; H3 (histamine H3)antagonist/inverse agonists, such as thioperamide,3-(1H-imidazol-4-yl)propyl N-(4-pentenyl)carbamate), clobenpropit,iodophenpropit, imoproxifan, GT2394 (Gliatech), and A331440,O-[3-(1H-imidazol-4-yl)propanol]carbamates (Kiec-Kononowicz, K. et al.,Pharmazie, 55:349-55 (2000)), piperidine-containing histamineH3-receptor antagonists (Lazewska, D. et al., Pharmazie, 56:927-32(2001), benzophenone derivatives and related compounds (Sasse, A. etal., Arch. Pharm. (Weinheim) 334:45-52 (2001)), substitutedN-phenylcarbamates (Reidemeister, S. et al., Pharmazie, 55:83-6 (2000)),and proxifan derivatives (Sasse, A. et al., J. Med. Chem. 43:3335-43(2000)) and histamine H3 receptor modulators such as those disclosed inWO02/15905, WO03/024928 and WO03/024929; leptin derivatives, such asthose disclosed in U.S. Pat. No. 5,552,524, U.S. Pat. No. 5,552,523,U.S. Pat. No. 5,552,522, U.S. Pat. No. 5,521,283, WO96/23513,WO96/23514, WO96/23515, WO96/23516, WO96/23517, WO96/23518, WO96/23519,and WO96/23520; leptin, including recombinant human leptin (PEG-OB,Hoffman La Roche) and recombinant methionyl human leptin (Amgen); lipaseinhibitors, such as tetrahydrolipstatin (orlistat/Xenical®), Triton WR1339, RHC80267, lipstatin, teasaponin, diethylumbelliferyl phosphate,FL-386, WAY-121898, Bay-N-3176, valilactone, esteracin, ebelactone A,ebelactone B, and RHC 80267, and those disclosed in patent publicationsWO01/77094, U.S. Pat. No. 4,598,089, U.S. Pat. No. 4,452,813,USUS5512565, U.S. Pat. No. 5,391,571, U.S. Pat. No. 5,602,151, U.S. Pat.No. 4,405,644, U.S. Pat. No. 4,189,438, and U.S. Pat. No. 4,242,453;lipid metabolism modulators such as maslinic acid, erythrodiol, ursolicacid uvaol, betulinic acid, betulin, and the like and compoundsdisclosed in WO03/011267; Mc4r (melanocortin 4 receptor) agonists, suchas CHIR86036 (Chiron), ME-10142, ME-10145, and HS-131 (Melacure), andthose disclosed in PCT publication Nos. WO99/64002, WO00/74679,WO01/991752, WO01/25192, WO01/52880, WO01/74844, WO01/70708, WO01/70337,WO01/91752, WO02/059095, WO02/059107, WO02/059108, WO02/059117,WO02/06276, WO02/12166, WO02/11715, WO02/12178, WO02/15909, WO02/38544,WO02/068387, WO02/068388, WO02/067869, WO02/081430, WO03/06604,WO03/007949, WO03/009847, WO03/009850, WO03/013509, and WO03/031410;Mc5r (melanocortin 5 receptor) modulators, such as those disclosed inWO97/19952, WO00/15826, WO00/15790, US20030092041; melanin-concentratinghormone 1 receptor (MCHR) antagonists, such as T-226296 (Takeda), SB568849, SNP-7941 (Synaptic), and those disclosed in patent publicationsWO01/21169, WO01/82925, WO01/87834, WO02/051809, WO02/06245,WO02/076929, WO02/076947, WO02/04433, WO02/51809, WO02/083134,WO02/094799, WO03/004027, WO03/13574, WO03/15769, WO03/028641,WO03/035624, WO03/033476, WO03/033480, JP13226269, and JP1437059; mGluR5modulators such as those disclosed in WO03/029210, WO03/047581,WO03/048137, WO03/051315, WO03/051833, WO03/053922, WO03/059904, and thelike; serotoninergic agents, such as fenfluramine (such as Pondimin®(Benzeneethanamine, N-ethyl-alpha-methyl-3-(trifluoromethyl)-,hydrochloride), Robbins), dexfenfluramine (such as Redux®(Benzeneethanamine, N-ethyl-alpha-methyl-3-(trifluoromethyl)-,hydrochloride), Interneuron) and sibutramine ((Meridia®,Knoll/Reductil™) including racemic mixtures, as optically pure isomers(+) and (−), and pharmaceutically acceptable salts, solvents, hydrates,clathrates and prodrugs thereof including sibutramine hydrochloridemonohydrate salts thereof, and those compounds disclosed in U.S. Pat.No. 4,746,680, U.S. Pat. No. 4,806,570, and U.S. Pat. No. 5,436,272,US20020006964, WO01/27068, and WO01/62341; NE (norepinephrine) transportinhibitors, such as GW 320659, despiramine, talsupram, and nomifensine;NPY 1 antagonists, such as BIBP3226, J-115814, BIBO 3304, LY-357897,CP-671906, GI-264879A, and those disclosed in U.S. Pat. No. 6,001,836,WO96/14307, WO01/23387, WO99/51600, WO01/85690, WO01/85098, WO01/85173,and WO01/89528; NPY5 (neuropeptide Y Y5) antagonists, such as 152,804,GW-569180A, GW-594884A, GW-587081X, GW-548118X, FR235208, FR226928,FR240662, FR252384, 1229U91, GI-264879A, CGP71683A, LY-377897,LY-366377, PD-160170, SR-120562A, SR-120819A, JCF-104, and H409/22 andthose compounds disclosed in patent publications U.S. Pat. No.6,140,354, U.S. Pat. No. 6,191,160, U.S. Pat. No. 6,218,408, U.S. Pat.No. 6,258,837, U.S. Pat. No. 6,313,298, U.S. Pat. No. 6,326,375, U.S.Pat. No. 6,329,395, U.S. Pat. No. 6,335,345, U.S. Pat. No. 6,337,332,U.S. Pat. No. 6,329,395, U.S. Pat. No. 6,340,683, EP01010691,EP-01044970, WO97/19682, WO97/20820, WO97/20821, WO97/20822, WO97/20823,WO98/27063, WO00/107409, WO00/185714, WO00/185730, WO00/64880,WO00/68197, WO00/69849, WO/0113917, WO01/09120, WO01/14376, WO01/85714,WO01/85730, WO01/07409, WO01/02379, WO01/23388, WO01/23389, WO01/44201,WO01/62737, WO01/62738, WO01/09120, WO02/20488, WO02/22592, WO02/48152,WO02/49648, WO02/051806, WO02/094789, WO03/009845, WO03/014083,WO03/022849, WO03/028726 and Norman et al, J. Med. Chem. 43:4288-4312(2000); opioid antagonists, such as nalmefene (REVEX®),3-methoxynaltrexone, methylnaltrexone, naloxone, and naltrexone (e.g.PT901; Pain Therapeutics, Inc.) and those disclosed in US20050004155 andWO00/21509; orexin antagonists, such as SB-334867-A and those disclosedin patent publications WO01/96302, WO01/68609, WO02/44172, WO02/51232,WO02/51838, WO02/089800, WO02/090355, WO03/023561, WO03/032991, andWO03/037847; PDE inhibitors (e.g. compounds which slow the degradationof cyclic AMP (cAMP) and/or cyclic GMP (cGMP) by inhibition of thephosphodiesterases, which can lead to a relative increase in theintracellular concentration of cAMP and cGMP; possible PDE inhibitorsare primarily those substances which are to be numbered among the classconsisting of the PDE3 inhibitors, the class consisting of the PDE4inhibitors and/or the class consisting of the PDE5 inhibitors, inparticular those substances which can be designated as mixed types ofPDE3/4 inhibitors or as mixed types of PDE3/4/5 inhibitors) such asthose disclosed in patent publications DE1470341, DE2108438, DE2123328,DE2305339, DE2305575, DE2315801, DE2402908, DE2413935, DE2451417,DE2459090, DE2646469, DE2727481, DE2825048, DE2837161, DE2845220,DE2847621, DE2934747, DE3021792, DE3038166, DE3044568, EP000718,EP0008408, EP0010759, EP0059948, EP0075436, EP0096517, EP01 12987, EP0116948, EP0150937, EP0158380, EP0161632, EP0161918, EP0167121, EP0199127,EP0220044, EP0247725, EP0258191, EP0272910, EP0272914, EP0294647,EP0300726, EP0335386, EP0357788, EP0389282, EP0406958, EP0426180,EP0428302, EP0435811, EP0470805, EP0482208, EP0490823, EP0506194,EP0511865, EP0527117, EP0626939, EP0664289, EP0671389, EP0685474,EP0685475, EP0685479, JP92234389, JP94329652, JP95010875, U.S. Pat. No.4,963,561, U.S. Pat. No. 5,141,931, WO9117991, WO9200968, WO9212961,WO9307146, WO9315044, WO9315045, WO9318024, WO9319068, WO9319720,WO9319747, WO9319749, WO9319751, WO9325517, WO9402465, WO9406423,WO9412461, WO9420455, WO9422852, WO9425437, WO9427947, WO9500516,WO9501980, WO9503794, WO9504045, WO9504046, WO9505386, WO9508534,WO9509623, WO9509624, WO9509627, WO9509836, WO9514667, WO9514680,WO9514681, WO9517392, WO9517399, WO9519362, WO9522520, WO9524381,WO9527692, WO9528926, WO9535281, WO9535282, WO9600218, WO9601825,WO9602541, WO9611917, DE3142982, DE1 116676, DE2162096, EP0293063,EP0463756, EP0482208, EP0579496, EP0667345 U.S. Pat. No. 6,331,543,US20050004222 (including those disclosed in formulas I-XIII andparagraphs 37-39, 85-0545 and 557-577), WO9307124, EP0163965, EP0393500,EP0510562, EP0553174, WO9501338 and WO9603399, as well as PDE5inhibitors (such as RX-RA-69, SCH-51866, KT-734, vesnarinone, zaprinast,SKF-96231, ER-21355, BF/GP-385, NM-702 and sildenafil (Viagra™)), PDE4inhibitors (such as etazolate, ICI63197, RP73401, imazolidinone(RO-20-1724), MEM 1414 (R1533/R1500; Pharmacia Roche), denbufylline,rolipram, oxagrelate, nitraquazone, Y-590, DH-6471, SKF-94120,motapizone, lixazinone, indolidan, olprinone, atizoram, KS-506-G,dipamfylline, BMY-43351, atizoram, arofylline, filaminast, PDB-093,UCB-29646, CDP-840, SKF-107806, piclamilast, RS-17597, RS-25344-000,SB-207499, TIBENELAST, SB-210667, SB-211572, SB-211600, SB-212066,SB-212179, GW-3600, CDP-840, mopidamol, anagrelide, ibudilast, amrinone,pimobendan, cilostazol, quazinone andN-(3,5-dichloropyrid-4-yl)-3-cyclopropylmethoxy4-difluoromethoxybenzamide,PDE3 inhibitors (such as ICI153, 100, bemorandane (RWJ 22867), MCI-154,UD-CG 212, sulmazole, ampizone, cilostamide, carbazeran, piroximone,imazodan, CI-930, siguazodan, adibendan, saterinone, SKF-95654,SDZ-MKS-492, 349-U-85, emoradan, EMD-53998, EMD-57033, NSP-306, NSP-307,revizinone, NM-702, WIN-62582 and WIN-63291, enoximone and milrinone,PDE3/4 inhibitors (such as benafentrine, trequinsin, ORG-30029,zardaverine, L-686398, SDZ-ISQ-844, ORG-20241, EMD-54622, andtolafentrine) and other PDE inhibitors (such as vinpocetin, papaverine,enprofylline, cilomilast, fenoximone, pentoxifylline, roflumilast,tadalafil (Cialis®), theophylline, and vardenafil (Levitra®);Neuropeptide Y2 (NPY2) agonists include but are not limited to:polypeptide YY and fragments and variants thereof (e.g. YY3-36(PYY3-36)(N. Engl. J. Med. 349:941, 2003; IKPEAPGE DASPEELNRY YASLRHYLNLVTRQRY (SEQ ID NO: 105)) and PYY agonists such as those disclosed inWO02/47712, WO03/026591, WO03/057235, and WO03/027637; serotoninreuptake inhibitors, such as, paroxetine, fluoxetine (Prozac™),fluvoxamine, sertraline, citalopram, and imipramine, and those disclosedin U.S. Pat. No. 6,162,805, U.S. Pat. No. 6,365,633, WO03/00663,WO01/27060, and WO01/162341; thyroid hormone β agonists, such as KB-2611(KaroBioBMS), and those disclosed in WO02/15845, WO97/21993, WO99/00353,GB98/284425, U.S. Provisional Application No. 60/183,223, and JapanesePatent Application No. JP 2000256190; UCP-I (uncoupling protein-1), 2,or 3 activators, such as phytanic acid, 4-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napthalenyl)-1-propenyl]benzoicacid (TTNPB), retinoic acid, and those disclosed in WO99/00123; β3 (betaadrenergic receptor 3) agonists, such as AJ9677/TAK677(Dainippon/Takeda), L750355 (Merck), CP331648 (Pfizer), CL-316,243, SB418790, BRL-37344, L-796568, BMS-196085, BRL-35135A, CGP12177A, BTA-243,GW 427353, Trecadrine, Zeneca D7114, N-5984 (Nisshin Kyorin), LY-377604(Lilly), SR 59119A, and those disclosed in U.S. Pat. No. 5,541,204, U.S.Pat. No. 5,770,615, U.S. Pat. No. 5,491,134, U.S. Pat. No. 5,776,983,US488064, U.S. Pat. No. 5,705,515, U.S. Pat. No. 5,451,677, WO94/18161,WO95/29159, WO97/46556, WO98/04526 and WO98/32753, WO01/74782,WO02/32897, WO03/014113, WO03/016276, WO03/016307, WO03/024948,WO03/024953 and WO03/037881; noradrenergic agents including, but notlimited to, diethylpropion (such as Tenuate® (1-propanone,2-(diethylamino)-1-phenyl-, hydrochloride), Merrell), dextroamphetamine(also known as dextroamphetamine sulfate, dexamphetamine, dexedrine,Dexampex, Femdex, Oxydess II, Robese, Spancap #1), mazindol ((or5-(p-chlorophenyl)-2,5-dihydro-3H-imidazo[2,1-a]isoindol-5-ol) such asSanorex®, Novartis or Mazanor®, Wyeth Ayerst), phenylpropanolamine (orBenzenemethanol, alpha-(1-aminoethyl)-, hydrochloride), phentermine ((orPhenol, 3-[[4,5-duhydro-1H-imidazol-2-yl)ethyl](4-methylpheny-1)amino],monohydrochloride) such as Adipex-P®, Lemmon, FASTIN®, Smith-KlineBeecham and Ionamin®, Medeva), phendimetrazine ((or(2S,3S)-3,4-Dimethyl-2phenylmorpholine L-(+)-tartrate (1:1)) such asMetra® (Forest), Plegine® (Wyeth-Ay erst), Prelu-2® (BoehringerIngelheim), and Statobex® (Lemmon), phendamine tartrate (such asThephorin®(2,3,4,9-Tetrahydro-2-methyl-9-phenyl-1H-indenol[2,1-c]pyridineL-(+)-tartrate (1:1)), Hoffmann-LaRoche), methamphetamine (such asDesoxyn®, Abbot ((S)—N, (alpha)-dimethylbenzeneethanaminehydrochloride)), and phendimetrazine tartrate (such as Bontril®Slow-Release Capsules, Amarin (-3,4-Dimethyl-2-phenylmorpholineTartrate); fatty acid oxidation upregulator/inducers such as Famoxin®(Genset); monamine oxidase inhibitors including but not limited tobefloxatone, moclobemide, brofaromine, phenoxathine, esuprone, befol,toloxatone, pirlindol, amiflamine, sercloremine, bazinaprine,lazabemide, milacemide, caroxazone and other certain compounds asdisclosed by WO01/12176; and other anti-obesity agents such as 5HT-2agonists, ACC (acetyl-CoA carboxylase) inhibitors such as thosedescribed in WO03/072197, alpha-lipoic acid (alpha-LA), AOD9604,appetite suppressants such as those in WO03/40107, ATL-962 (AlizymePLC), benzocaine, benzphetamine hydrochloride (Didrex), bladderwrack(focus vesiculosus), BRS3 (bombesin receptor subtype 3) agonists,bupropion, caffeine, CCK agonists, chitosan, chromium, conjugatedlinoleic acid, corticotropin-releasing hormone agonists,dehydroepiandrosterone, DGAT1 (diacylglycerol acyltransferase 1)inhibitors, DGAT2 (diacylglycerol acyltransferase 2) inhibitors,dicarboxylate transporter inhibitors, ephedra, exendin-4 (an inhibitorof glp-1) FAS (fatty acid synthase) inhibitors (such as Cerulenin andC75), fat resorption inhibitors (such as those in WO03/053451, and thelike), fatty acid transporter inhibitors, natural water soluble fibers(such as psyllium, plantago, guar, oat, pectin), galanin antagonists,galega (Goat's Rue, French Lilac), garcinia cambogia, germander(teucrium chamaedrys), ghrelin antibodies and ghrelin antagonists (suchas those disclosed in WO01/87335, and WO02/08250), polypeptide hormonesand variants thereof which affect the islet cell secretion, such as thehormones of the secretin/gastric inhibitory polypeptide (GIP)/vasoactiveintestinal polypeptide (VIP)/pituitary adenylate cyclase activatingpolypeptide (PACAP)/glucagon-like polypeptide II(GLP-II)/glicentin/glucagon gene family and/or those of theadrenomedullin/amylin/calcitonin gene related polypeptide (CGRP) genefamily includingGLP-1 (glucagon-like polypeptide 1) agonists (e.g. (1)exendin-4, (2) those GLP-I molecules described in US20050130891including GLP-1(7-34), GLP-1(7-35), GLP-1(7-36) or GLP-1(7-37) in itsC-terminally carboxylated or amidated form or as modified GLP-Ipolypeptides and modifications thereof including those described inparagraphs 17-44 of US20050130891, and derivatives derived fromGLP-1-(7-34)COOH and the corresponding acid amide are employed whichhave the following general formula:R—NH-HAEGTFTSDVSYLEGQAAKEFIAWLVK-CONH₂ wherein R═H or an organiccompound having from 1 to 10 carbon atoms (SEQ ID NO: 106). Preferably,R is the residue of a carboxylic acid. Particularly preferred are thefollowing carboxylic acid residues: formyl, acetyl, propionyl,isopropionyl, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl,tert-butyl.) and glp-1 (glucagon-like polypeptide-1), glucocorticoidantagonists, glucose transporter inhibitors, growth hormonesecretagogues (such as those disclosed and specifically described inU.S. Pat. No. 5,536,716), interleukin-6 (IL-6) and modulators thereof(as in WO03/057237, and the like), L-carnitine, Mc3r (melanocortin 3receptor) agonists, MCH2R (melanin concentrating hormone 2R)agonist/antagonists, melanin concentrating hormone antagonists,melanocortin agonists (such as Melanotan II or those described in WO99/64002 and WO 00/74679), nomame herba, phosphate transporterinhibitors, phytopharm compound 57 (CP 644,673), pyruvate, SCD-I(stearoyl-CoA desaturase-1) inhibitors, T71 (Tularik, Inc., BoulderColo.), Topiramate (Topimax®, indicated as an anti-convulsant which hasbeen shown to increase weight loss), transcription factor modulators(such as those disclosed in WO03/026576), β-hydroxy steroiddehydrogenase-1 inhibitors (β-HSD-I), β-hydroxy-β-methylbutyrate, p57(Pfizer), Zonisamide (Zonegran™, indicated as an anti-epileptic whichhas been shown to lead to weight loss), and the agents disclosed inUS20030119428 paragraphs 20-26.

Anti-Diabetic Agents

The Aad-GCRA peptides described herein can be used in therapeuticcombination with one or more anti-diabetic agents, including but notlimited to: PPARγ agonists such as glitazones (e.g., WAY-120,744, AD5075, balaglitazone, ciglitazone, darglitazone (CP-86325, Pfizer),englitazone (CP-68722, Pfizer), isaglitazone (MIT/J&J), MCC-555(Mitsibishi disclosed in U.S. Pat. No. 5,594,016), pioglitazone (such assuch as Actos™ pioglitazone; Takeda), rosiglitazone (Avandia™; SmithKline Beecham), rosiglitazone maleate, troglitazone (Rezulin®, disclosedin U.S. Pat. No. 4,572,912), rivoglitazone (CS-O1 1, Sankyo), GL-262570(Glaxo Welcome), BRL49653 (disclosed in WO98/05331), CLX-0921, 5-BTZD,GW-0207, LG-100641, JJT-501 (JPNT/P&U), L-895645 (Merck), R-119702(Sankyo/Pfizer), NN-2344 (Dr. Reddy/NN), YM-440 (Yamanouchi), LY-300512,LY-519818, R483 (Roche), T131 (Tularik), and the like and compoundsdisclosed in U.S. Pat. No. 4,687,777, U.S. Pat. No. 5,002,953, U.S. Pat.No. 5,741,803, U.S. Pat. No. 5,965,584, U.S. Pat. No. 6,150,383, U.S.Pat. No. 6,150,384, U.S. Pat. No. 6,166,042, U.S. Pat. No. 6,166,043,U.S. Pat. No. 6,172,090, U.S. Pat. No. 6,211,205, U.S. Pat. No.6,271,243, U.S. Pat. No. 6,288,095, U.S. Pat. No. 6,303,640, U.S. Pat.No. 6,329,404, U.S. Pat. No. 5,994,554, WO97/10813, WO97/27857,WO97/28115, WO97/28137, WO97/27847, WO00/76488, WO03/000685,WO03/027112, WO03/035602, WO03/048130, WO03/055867, and pharmaceuticallyacceptable salts thereof; biguanides such as metformin hydrochloride(N,N-dimethylimidodicarbonimidic diamide hydrochloride, such asGlucophage™, Bristol-Myers Squibb); metformin hydrochloride withglyburide, such as Glucovance™, Bristol-Myers Squibb); buformin(Imidodicarbonimidic diamide, N-butyl-); etoformine(1-Butyl-2-ethylbiguanide, Schering A. G.); other metformin salt forms(including where the salt is chosen from the group of, acetate,benzoate, citrate, ftimarate, embonate, chlorophenoxyacetate, glycolate,palmoate, aspartate, methanesulphonate, maleate,parachlorophenoxyisobutyrate, formate, lactate, succinate, sulphate,tartrate, cyclohexanecarboxylate, hexanoate, octanoate, decanoate,hexadecanoate, octodecanoate, benzenesulphonate, trimethoxybenzoate,paratoluenesulphonate, adamantanecarboxylate, glycoxylate, glutamate,pyrrolidonecarboxylate, naphthalenesulphonate, 1-glucosephosphate,nitrate, sulphite, dithionate and phosphate), and phenformin; proteintyrosine phosphatase-IB (PTP-IB) inhibitors, such as A-401,674, KR61639, OC-060062, OC-83839, OC-297962, MC52445, MC52453, ISIS 113715,and those disclosed in WO99/585521, WO99/58518, WO99/58522, WO99/61435,WO03/032916, WO03/032982, WO03/041729, WO03/055883, WO02/26707,WO02/26743, JP2002114768, and pharmaceutically acceptable salts andesters thereof; sulfonylureas such as acetohexamide (e.g. Dymelor, EliLilly), carbutamide, chlorpropamide (e.g. Diabinese®, Pfizer),gliamilide (Pfizer), gliclazide (e.g. Diamcron, Servier Canada Inc),glimepiride (e.g. disclosed in U.S. Pat. No. 4,379,785, such as Amaryl,Aventis), glipentide, glipizide (e.g. Glucotrol or Glucotrol XL ExtendedRelease, Pfizer), gliquidone, glisolamide, glyburide/glibenclamide (e.g.Micronase or Glynase Prestab, Pharmacia & Upjohn and Diabeta, Aventis),tolazamide (e.g. Tolinase), and tolbutamide (e.g. Orinase), andpharmaceutically acceptable salts and esters thereof; meglitinides suchas repaglinide (e.g. Pranidin®, Novo Nordisk), KAD 1229 (PF/Kissei), andnateglinide (e.g. Starlix®, Novartis), and pharmaceutically acceptablesalts and esters thereof; a glucoside hydrolase inhibitors (or glucosideinhibitors) such as acarbose (e.g. Precose™, Bayer disclosed in U.S.Pat. No. 4,904,769), miglitol (such as GLYSET™, Pharmacia & Upjohndisclosed in U.S. Pat. No. 4,639,436), camiglibose (Methyl6-deoxy-6-[(2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidino]-alpha-D-glucopyranoside,Marion Merrell Dow), voglibose (Takeda), adiposine, emiglitate,pradimicin-Q, salbostatin, CKD-711, MDL-25,637, MDL-73,945, and MOR 14,and the compounds disclosed in U.S. Pat. No. 4,062,950, U.S. Pat. No.4,174,439, U.S. Pat. No. 4,254,256, U.S. Pat. No. 4,701,559, U.S. Pat.No. 4,639,436, U.S. Pat. No. 5,192,772, U.S. Pat. No. 4,634,765, U.S.Pat. No. 5,157,116, U.S. Pat. No. 5,504,078, U.S. Pat. No. 5,091,418,U.S. Pat. No. 5,217,877, and WO01/47528 (polyamines); α-amylaseinhibitors such as tendamistat, trestatin, and A1-3688, and thecompounds disclosed in U.S. Pat. No. 4,451,455, U.S. Pat. No. 4,623,714,and U.S. Pat. No. 4,273,765; SGLT2 inhibtors including those disclosedin U.S. Pat. No. 6,414,126 and U.S. Pat. No. 6,515,117; an aP2 inhibitorsuch as disclosed in U.S. Pat. No. 6,548,529; insulin secreatagoguessuch as linogliride, A-4166, forskilin, dibutyrl cAMP,isobutylmethylxanthine (IBMX), and pharmaceutically acceptable salts andesters thereof; fatty acid oxidation inhibitors, such as clomoxir, andetomoxir, and pharmaceutically acceptable salts and esters thereof; A2antagonists, such as midaglizole, isaglidole, deriglidole, idazoxan,earoxan, and fluparoxan, and pharmaceutically acceptable salts andesters thereof; insulin and related compounds (e.g. insulin mimetics)such as biota, LP-100, novarapid, insulin detemir, insulin lispro,insulin glargine, insulin zinc suspension (lente and ultralente),Lys-Pro insulin, GLP-I (1-36) amide, GLP-I (73-7) (insulintropin,disclosed in U.S. Pat. No. 5,614,492), LY-315902 (Lilly), GLP-I(7-36)-NH2), AL-401 (Autoimmune), certain compositions as disclosed inU.S. Pat. No. 4,579,730, U.S. Pat. No. 4,849,405, U.S. Pat. No.4,963,526, U.S. Pat. No. 5,642,868, U.S. Pat. No. 5,763,396, U.S. Pat.No. 5,824,638, U.S. Pat. No. 5,843,866, U.S. Pat. No. 6,153,632, U.S.Pat. No. 6,191,105, and WO 85/05029, and primate, rodent, or rabbitinsulin including biologically active variants thereof including allelicvariants, more preferably human insulin available in recombinant form(sources of human insulin include pharmaceutically acceptable andsterile formulations such as those available from Eli Lilly(Indianapolis, Ind. 46285) as Humulin™ (human insulin rDNA origin), alsosee the THE PHYSICIAN'S DESK REFERENCE, 55.sup.th Ed. (2001) MedicalEconomics, Thomson Healthcare (disclosing other suitable humaninsulins); non-thiazolidinediones such as JT-501 and farglitazar(GW-2570/GI-262579), and pharmaceutically acceptable salts and estersthereof; PPARα/γ dual agonists such as AR-HO39242 (Aztrazeneca),GW-409544 (Glaxo-Wellcome), BVT-142, CLX-0940, GW-1536, GW-1929,GW-2433, KRP-297 (Kyorin Merck; 5-[(2,4-Dioxo thiazolidinyl)methyl]methoxy-N-[[4-(trifluoromethyl)phenyl] methyljbenzamide), L-796449,LR-90, MK-0767 (Merck/Kyorin/Banyu), SB 219994, muraglitazar (BMS),tesaglitzar (Astrazeneca), reglitazar (JTT-501) and those disclosed inWO99/16758, WO99/19313, WO99/20614, WO99/38850, WO00/23415, WO00/23417,WO00/23445, WO00/50414, WO01/00579, WO01/79150, WO02/062799,WO03/004458, WO03/016265, WO03/018010, WO03/033481, WO03/033450,WO03/033453, WO03/043985, WO 031053976, U.S. application Ser. No.09/664,598, filed Sep. 18, 2000, Murakami et al. Diabetes 47, 1841-1847(1998), and pharmaceutically acceptable salts and esters thereof; otherinsulin sensitizing drugs; VPAC2 receptor agonists; GLK modulators, suchas those disclosed in WO03/015774; retinoid modulators such as thosedisclosed in WO03/000249; GSK 30/GSK 3 inhibitors such as4-[2-(2-bromophenyl)-4-(4-fluorophenyl-1H-imidazol-5-yl]pyridine andthose compounds disclosed in WO03/024447, WO03/037869, WO03/037877,WO03/037891, WO03/068773, EP1295884, EP1295885, and the like; glycogenphosphorylase (HGLPa) inhibitors such as CP-368,296, CP-316,819,BAYR3401, and compounds disclosed in WO01/94300, WO02/20530,WO03/037864, and pharmaceutically acceptable salts or esters thereof;ATP consumption promotors such as those disclosed in WO03/007990; TRB3inhibitors; vanilloid receptor ligands such as those disclosed inWO03/049702; hypoglycemic agents such as those disclosed in WO03/015781and WO03/040114; glycogen synthase kinase 3 inhibitors such as thosedisclosed in WO03/035663 agents such as those disclosed in WO99/51225,US20030134890, WO01/24786, and WO03/059870; insulin-responsive DNAbinding protein-1 (IRDBP-I) as disclosed in WO03/057827, and the like;adenosine A2 antagonists such as those disclosed in WO03/035639,WO03/035640, and the like; PPARδ agonists such as GW 501516, GW 590735,and compounds disclosed in JP10237049 and WO02/14291; dipeptidylpeptidase IV (DP-IV) inhibitors, such as isoleucine thiazolidide,NVP-DPP728A(1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(S)-pyrrolidine,disclosed by Hughes et al, Biochemistry, 38(36), 11597-11603, 1999),P32/98, NVP-LAF-237, P3298, TSL225(tryptophyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid, disclosedby Yamada et al, Bioorg. & Med. Chem. Lett. 8 (1998) 1537-1540), valinepyrrolidide, TMC-2A/2B/2C, CD-26 inhibitors, FE999011, P9310/K364, VIP0177, DPP4, SDZ 274-444, 2-cyanopyrrolidides and 4-cyanopyrrolidides asdisclosed by Ashworth et al, Bioorg. & Med. Chem. Lett., Vol. 6, No. 22,pp 1163-1166 and 2745-2748 (1996), and the compounds disclosed in U.S.Pat. No. 6,395,767, U.S. Pat. No. 6,573,287, U.S. Pat. No. 6,395,767(compounds disclosed include BMS-477118, BMS-471211 and BMS 538,305),WO99/38501, WO99/46272, WO99/67279, WO99/67278, WO99/61431 WO03/004498,WO03/004496, EP1258476, WO02/083128, WO02/062764, WO03/000250,WO03/002530, WO03/002531, WO03/002553, WO03/002593, WO03/000180, andWO03/000181; GLP-I agonists such as exendin-3 and exendin-4 (includingthe 39 aa polypeptide synthetic exendin-4 called Exenatide®), andcompounds disclosed in US2003087821 and NZ 504256, and pharmaceuticallyacceptable salts and esters thereof; peptides including amlintide andSymlin® (pramlintide acetate); and glycokinase activators such as thosedisclosed in US2002103199 (fused heteroaromatic compounds) andWO02/48106 (isoindolin-1-one-substituted propionamide compounds).

Phosphodiesterase Inhibitors

The Aad-GCRA peptides described herein can be used in combinationtherapy with a phosphodiesterase inhibitor. PDE inhibitors are thosecompounds which slow the degradation of cyclic AMP (cAMP) and/or cyclicGMP (cGMP) by inhibition of the phosphodiesterases, which can lead to arelative increase in the intracellular concentration of c AMP and/orcGMP. Possible PDE inhibitors are primarily those substances which areto be numbered among the class consisting of the PDE3 inhibitors, theclass consisting of the PDE4 inhibitors and/or the class consisting ofthe PDE5 inhibitors, in particular those substances which can bedesignated as mixed types of PDE3/4 inhibitors or as mixed types ofPDE3/4/5 inhibitors. By way of example, those PDE inhibitors may bementioned such as are described and/or claimed in the following patentapplications and patents: DE1470341, DE2108438, DE2123328, DE2305339,DE2305575, DE2315801, DE2402908, DE2413935, DE2451417, DE2459090,DE2646469, DE2727481, DE2825048, DE2837161, DE2845220, DE2847621,DE2934747, DE3021792, DE3038166, DE3044568, EP000718, EP0008408,EP0010759, EP0059948, EP0075436, EP0096517, EPO1 12987, EPO1 16948,EP0150937, EP0158380, EP0161632, EP0161918, EP0167121, EP0199127,EP0220044, EP0247725, EP0258191, EP0272910, EP0272914, EP0294647,EP0300726, EP0335386, EP0357788, EP0389282, EP0406958, EP0426180,EP0428302, EP0435811, EP0470805, EP0482208, EP0490823, EP0506194,EP0511865, EP0527117, EP0626939, EP0664289, EP0671389, EP0685474,EP0685475, EP0685479, JP92234389, JP94329652, JP95010875, U.S. Pat. Nos.4,963,561, 5,141,931, WO9117991, WO9200968, WO9212961, WO9307146,WO9315044, WO9315045, WO9318024, WO9319068, WO9319720, WO9319747,WO9319749, WO9319751, WO9325517, WO9402465, WO9406423, WO9412461,WO9420455, WO9422852, WO9425437, WO9427947, WO9500516, WO9501980,WO9503794, WO9504045, WO9504046, WO9505386, WO9508534, WO9509623,WO9509624, WO9509627, WO9509836, WO9514667, WO9514680, WO9514681,WO9517392, WO9517399, WO9519362, WO9522520, WO9524381, WO9527692,WO9528926, WO9535281, WO9535282, WO9600218, WO9601825, WO9602541,WO9611917, DE3142982, DE1 116676, DE2162096, EP0293063, EP0463756,EP0482208, EP0579496, EP0667345 U.S. Pat. No. 6,331,543, US20050004222(including those disclosed in formulas I-XIII and paragraphs 37-39,85-0545 and 557-577) and WO9307124, EP0163965, EP0393500, EP0510562,EP0553174, WO9501338 and WO9603399. PDE5 inhibitors which may bementioned by way of example are RX-RA-69, SCH-51866, KT-734,vesnarinone, zaprinast, SKF-96231, ER-21355, BF/GP-385, NM-702 andsildenafil (Viagra®). PDE4 inhibitors which may be mentioned by way ofexample are RO-20-1724, MEM 1414 (R1533/R1500; Pharmacia Roche),DENBUFYLLINE, ROLIPRAM, OXAGRELATE, NITRAQUAZONE, Y-590, DH-6471,SKF-94120, MOTAPIZONE, LIXAZINONE, INDOLIDAN, OLPRINONE, ATIZORAM,KS-506-G, DIPAMFYLLINE, BMY-43351, ATIZORAM, AROFYLLINE, FILAMINAST,PDB-093, UCB-29646, CDP-840, SKF-107806, PICLAMILAST, RS-17597,RS-25344-000, SB-207499, TIBENELAST, SB-210667, SB-211572, SB-211600,SB-212066, SB-212179, GW-3600, CDP-840, MOPIDAMOL, ANAGRELIDE,IBUDILAST, AMRINONE, PIMOBENDAN, CILOSTAZOL, QUAZINONE andN-(3,5-dichloropyrid-4-yl)-3-cyclopropylmethoxy4-difluoromethoxybenzamide.PDE3 inhibitors which may be mentioned by way of example are SULMAZOLE,AMPIZONE, CILOSTAMIDE, CARBAZERAN, PIROXIMONE, IMAZODAN, CI-930,SIGUAZODAN, ADIBENDAN, SATERINONE, SKF-95654, SDZ-MKS-492, 349-U-85,EMORADAN, EMD-53998, EMD-57033, NSP-306, NSP-307, REVIZINONE, NM-702,WIN-62582 and WIN-63291, ENOXIMONE and MILRINONE. PDE3/4 inhibitorswhich may be mentioned by way of example are BENAFENTRINE, TREQUINSIN,ORG-30029, ZARDAVERINE, L-686398, SDZ-ISQ-844, ORG-20241, EMD-54622, andTOLAFENTRINE. Other PDE inhibitors include: cilomilast, pentoxifylline,roflumilast, tadalafil (Cialis®), theophylline, and vardenafil(Levitra®), zaprinast (PDE5 specific).

Anti-Uterine Contractions Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy (for example, in order to decrease or inhibit uterinecontractions) with a tocolytic agent including but not limited tobeta-adrenergic agents, magnesium sulfate, prostaglandin inhibitors, andcalcium channel blockers.

Anti-Neoplastic Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy with an antineoplastic agents including but not limited toalkylating agents, epipodophyllotoxins, nitrosoureas, antimetabolites,vinca alkaloids, anthracycline antibiotics, nitrogen mustard agents, andthe like. Particular anti-neoplastic agents may include tamoxifen,taxol, etoposide and 5-fluorouracil.

The Aad-GCRA peptides described herein can be used in combinationtherapy (for example as in a chemotherapeutic composition) with anantiviral and monoclonal antibody therapies.

Agents to Treat Congestive Heart Failure

The Aad-GCRA peptides described herein can be used in combinationtherapy (for example, in prevention/treatment of congestive heartfailure or another method described herein) with the partial agonist ofthe nociceptin receptor ORL1 described by Dooley et al. (The Journal ofPharmacology and Experimental Therapeutics, 283 (2): 735-741, 1997). Theagonist is a hexapeptide having the amino acid sequence Ac—RYY (RK) (WI)(RK)—NH2 (“the Dooley polypeptide”), where the brackets show allowablevariation of amino acid residue. Thus Dooley polypeptide can include butare not limited to KYYRWR (SEQ ID NO: 107), RYYRWR (SEQ ID NO: 108),KWRYYR (SEQ ID NO: 109), RYYRWK (SEQ ID NO: 110), RYYRWK (all-D aminacids) (SEQ ID NO: 111), RYYRIK (SEQ ID NO: 112), RYYRIR (SEQ ID NO:113), RYYKIK (SEQ ID NO: 114), RYYKIR (SEQ ID NO: 115), RYYKWR (SEQ IDNO: 116), RYYKWK (SEQ ID NO: 117), and KYYRWK (SEQ ID NO: 118), whereinthe amino acid residues are in the L-form unless otherwise specified.The Aad-GCRA peptides described herein can also be used in combinationtherapy with polypeptide conjugate modifications of the Dooleypolypeptide described in WO0198324.

Fibrate

The Aad-GCRA peptides described herein can be used in combinationtherapy with a fibrate. The term “fibrate” is also interchangeably usedherein and in the art with the term “fibric acid derivative,” and meansany of the fibric acid derivatives useful in the methods describedherein, e.g., fenofibrate. Fenofibrate is a fibrate compound, otherexamples of which include, for example, bezafibrate, beclofibrate,benzafibrate, binifibrate, ciprofibrate, clinofibrate, clofibrate,etofibrate, gemcabene, gemfibrozil, lifibrol, nicofibrate, pirifibrate,ronifibrate, simfibrate, theofibrate, etc.

Lipid Altering Agents

The Aad-GCRA peptides described herein can be used in combinationtherapy with a lipid altering agent. As used herein the term “lipidaltering agent” or “dyslipidemia agent” refers to compounds including,but not limited to, bile acid sequestrants such as cholestyramine (astyrene-divinylbenzene copolymer containing quaternary ammonium cationicgroups capable of binding bile acids, such as QUESTRAN® or QUESTRANLIGHT® cholestyramine which are available from Bristol-Myers Squibb),colesevelam hydrochloride (such as WELCHOL® Tablets (polyallylaminehydrochloride) cross-linked with epichlorohydrin and alkylated with1-bromodecane and (6-bromohexyl)-trimethylammonium bromide) which areavailable from Sankyo), colestipol (a copolymer of diethylenetriamineand 1-chloro-2,3-epoxypropane, such as COLESTID® tablets which areavailable from Pharmacia), dialkylaminoalkyl derivatives of across-linked dextran, LOCHOLEST®, DEAE-Sephadex (SECHOLEX®,POLICEXIDE®), water soluble derivatives such as 3,3-ioene,N-(cycloalkyl)alkylamines and poliglusam, insoluble quaternizedpolystyrenes, saponins and mixtures thereof and those bile acidsequestrants disclosed in WO97/11345, WO98/57652, U.S. Pat. No.3,692,895, and U.S. Pat. No. 5,703,188. Suitable inorganic cholesterolsequestrants include bismuth salicylate plus montmorillonite clay,aluminum hydroxide and calcium carbonate antacids.

HMG-CoA Reductase Inhibitors

The Aad-GCRA peptides described herein can be used in combinationtherapy with a HMG-CoA reductase inhibitor. HMG-CoA reductase inhibitorsare dyslipidemic agents that can be used in therapeutic combinationswith compounds described herein. Suitable HMG-CoA reductase inhibitorsfor use in therapeutic combination with a compounds described hereininclude: atorvastatin (LIPITOR®; disclosed in U.S. Pat. No. 4,681,893,U.S. Pat. No. 5,385,929 and U.S. Pat. No. 5,686,104), atorvastatincalcium (disclosed in U.S. Pat. No. 5,273,995), dihydrocompactin,(disclosed in U.S. Pat. No. 4,450,171), bervastatin (disclosed in U.S.Pat. No. 5,082,859), carvastatin, cerivastatin (BAYCOL®; disclosed inU.S. Pat. No. 5,006,530, U.S. Pat. No. 5,502,199, and U.S. Pat. No.5,177,080), crilvastatin, dalvastatin (disclosed in EP738510A2),fluvastatin (LESCOL®; disclosed in U.S. Pat. No. 4,739,073 andUS534772), glenvastatin, fluindostatin (disclosed in EP363934A1),velostatin (visinolin; disclosed in U.S. Pat. No. 4,448,784 and U.S.Pat. No. 4,450,171), lovastatin (mevinolin; MEVACOR® (Merck and Co.) andrelated compounds disclosed in U.S. Pat. No. 4,231,938), mevastatin (andrelated compound disclosed in U.S. Pat. No. 3,983,140), compactin (andrelated compounds disclosed in U.S. Pat. No. 4,804,770), pravastatin(also known as NK-104, itavastatin, nisvastatin, nisbastatin disclosedin U.S. Pat. No. 5,102,888), pravastatin (PRAVACHOL® (Bristol MyersSquibb) and related compounds disclosed in U.S. Pat. No. 4,346,227),rivastatin (sodium7-(4-fluorophenyl)-2,6-diisopropyl-5-methoxymethylpyridin-3-yl)-3,5-dihydroxy-6-heptanoate),rosuvastatin (CRESTOR®; also known as ZD-4522 disclosed in U.S. Pat. No.5,260,440), atavastatin, visastatin, simvastatin (ZOCOR® (Merck and Co.)and related compounds as disclosed in U.S. Pat. No. 4,448,784 and U.S.Pat. No. 4,450,171), simvastatin, CI-981, compounds disclosed inWO03/033481, U.S. Pat. No. 4,231,938, U.S. Pat. No. 4,444,784, U.S. Pat.No. 4,647,576, U.S. Pat. No. 4,686,237, U.S. Pat. No. 4,499,289, U.S.Pat. No. 4,346,227, U.S. Pat. No. 5,753,675, U.S. Pat. No. 4,613,610,EP0221025, and EP491226, and optical or geometric isomers thereof; andnontoxic pharmaceutically acceptable salts, N-oxides, esters, quaternaryammonium salts, and prodrugs thereof. In HMG-CoA reductase inhibitorswhere an open-acid form can exist, salt and ester forms may preferablybe formed from the open-acid, and all such forms are included within themeaning of the term “HMG-CoA reductase inhibitor” as used herein.Pharmaceutically acceptable salts with respect to the HMG-CoA reductaseinhibitor includes non-toxic salts of the compounds which are generallyprepared by reacting the free acid with a suitable organic or inorganicbase, particularly those formed from cations such as sodium, potassium,aluminum, calcium, lithium, magnesium, zinc and tetramethylammonium, aswell as those salts formed from amines such as ammonia, ethylenediamine,N-methylglucamine, lysine, arginine, ornithine (Orn), choline,N,N′-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine,N-benzylphenethylamine,1-p-chlorobenzyl-2-pyrrolidine-1′-yl-methylbenzim-idazole, diethylamine,piperazine, and tris(hydroxymethyl) aminomethane. Further examples ofsalt forms of HMG-CoA reductase inhibitors may include, but are notlimited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate,bitartrate, borate, bromide, calcium edetate, camsylate, carbonate,chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate,estolate, esylate, fumarate, gluceptate, gluconate, glutamate,glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,hydrochloride, hydroxynapthoate, iodide, isothionate, lactate,lactobionate, laurate, malate, maleate, mandelate, mesylate,methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote,palmitate, pantothenate, phosphate/diphosphate, polygalacturonate,salicylate, stearate, subacetate, succinate, tannate, tartrate,teoclate, tosylate, triethiodide, and valerate.

Other dyslipidemic agents which can be used in therapeutic combinationwith a compound described herein include: HMG-CoA synthase inhibitorssuch as L-659,699 ((E E)-I1-[3′R-(hydroxy-methyl)-4′-oxo-2′R-oxetanyl]-3,5,7R-trimethyl-2,4-undecadienoicacid) and those disclosed in U.S. Pat. No. 5,120,729, U.S. Pat. No.5,064,856, and U.S. Pat. No. 4,847,271; cholesterol absorptioninhibitors such as plant sterols, plant stanols and/or fatty acidestesrs of plant stanols such as sitostanol ester used in BENECOL®margarine, stanol esters, beta-sitosterol, and sterol glycosides such astiqueside. Other cholesterol absorption inhibitors include1,4-Diphenylazetidin-2-ones; 4-biarylyl-1-phenylazetidin-2-ones;4-(hydroxyphenyl)azetidin-2-ones;1,4-diphenyl-3-hydroxyalkyl-2-azetidinones;4-biphenyl-1-phenylazetidin-2-ones; 4-biarylyl-1-phenylazetidin-2-ones;and 4-biphenylylazetidinones.acyl coenzyme A-cholesterol acyltransferase (ACAT) inhibitors such as avasimibe (Current Opinion inInvestigational Drugs. 3(9):291-297 (2003)), eflucimibe, HL-004,lecimibe, DuP-128, KY505, SMP 797, CL-277,082 (Clin Pharmacol Ther.48(2): 189-94 (1990)) and the like; and those disclosed in US55 10379,WO96/26948 and WO96/10559; CETP inhibitors such as JTT 705 identified asin Nature 406, (6792):203-7 (2000), torcetrapib (CP-529,414 described inUS20030186952 and WO00/017164), CP 532,632, BAY63-2149, SC 591, SC 795,and the like including those described in Current Opinion inInvestigational Drugs. 4(3):291-297 (2003) and those disclosed in J.Antibiot, 49(8): 815-816 (1996), and Bioorg. Med. Chem. Lett,6:1951-1954 (1996) and patent publications US55 12548, U.S. Pat. No.6,147,090, WO99/20302, WO99/14204, WO99/41237, WO95/04755, WO96/15141,WO96/05227, WO038721, EP796846, EP818197, EP818448, DE19704244,DE19741051, DE19741399, DE197042437, DE19709125, DE19627430, DE19832159,DE19741400, JP 11049743, and JP 09059155; squalene synthetase inhibitorssuch as squalestatin-1, TAK-475, and those disclosed in U.S. Pat. No.4,871,721, U.S. Pat. No. 4,924,024, US57 12396 (α-phosphono-sulfonates),Biller et al (1988) J. Med. Chem., 31:1869 (e.g. isoprenoid(phosphinyl-methyl)phosphonates), Biller et al (1996) CurrentPharmaceutical Design, 2:1, P. Ortiz de Montellano et al (1977) J. Med.Chem. 20:243 (terpenoid pyrophosphates), Corey and Volante (1976) J. Am.Chem. Soc, 98:1291 (famesyl diphosphate analog A and presqualenepyrophosphate (PSQ-PP) analogs), McClard et al (1987) J.A.C.S., 109:5544(phosphinylphosphonates), Capson, T. L., PhD dissertation, June, 1987,Dept. Med. Chem. U of Utah, Abstract, Table of Contents, pp 16, 17,40-43, 48-51, Summary, (cyclopropanes), Curr. Op. Ther. Patents (1993)861, and patent publications EP0567026A1, EP0645378A1, EP0645377A1,EP0611749A1, EP0705607A2, EP0701725A1, and WO96/09827; antioxidants suchas probucol (and related compounds disclosed in U.S. Pat. No.3,674,836), probucol derivatives such as AGI-1067 (and other derivativesdisclosed in U.S. Pat. No. 6,121,319 and U.S. Pat. No. 6,147,250),tocopherol, ascorbic acid, 3-carotene, selenium and vitamins such asvitamin B6 or vitamin B12 and pharmaceutically acceptable salts andesters thereof; PPARα agonists such as those disclosed in U.S. Pat. No.6,028,109 (fluorophenyl compounds), WO00/75103 (substitutedphenylpropionic compounds), WO98/43081 and fibric acid derivatives(fibrates) such as beclofibrate, benzafibrate, bezafibrate (C.A.S.Registry No. 41859-67-0, see U.S. Pat. No. 3,781,328), binifibrate(C.A.S. Registry No. 69047-39-8, see BE884722), ciprofibrate (C.A.S.Registry No. 52214-84-3, see U.S. Pat. No. 3,948,973), clinofibrate(C.A.S. Registry No. 30299-08-2, see U.S. Pat. No. 3,716,583),clofibrate (such as ethyl 2-(p-chlorophenoxy)-2-methyl-propionate, e.g.Atromid-S® capsules (Wyeth-Ayerst), etofibrate, fenofibrate (such asTricor® micronized fenofibrate((2-[4-(4-chlorobenzoyl)phenoxy]-2-methyl-propanoic acid, 1-methylethylester; Abbott Laboratories) or Lipanthyl® micronized fenofibrate(Labortoire Founier, France)), gemcabene, gemfibrozil (such as5-(2,5-dimethylphenoxy)-2,2-dimethylpentanoic acid, e.g. Lopid® tablets(Parke Davis)), lifibrol, GW 7647, BM 170744, LY5 18674 and thosefibrate and fibrate acid derivatives disclosed in WO03/033456,WO03/033481, WO03/043997, WO03/048116, WO03/053974, WO03/059864, andWO03/05875; FXR receptor modulators such as GW 4064, SR 103912, and thelike; LXR receptor modulators such as GW 3965, T9013137, and XTC0179628,and those disclosed in US20030125357, WO03/045382, WO03/053352,WO03/059874, and the like; HM74 and HM74A (human HM74A is GenbankAccession No. AY148884 and rat HM74A is EMM_patAR09 8624) receptoragonists such as nicotinic acid (niacin) and derivatives thereof (e.g.compounds comprising a pyridine-3-carboxylate structure or apyrazine-2-carboxylate structure, including acid forms, salts, esters,zwitterions and tautomers, where available) including but not limited tothose disclosed in Wise et al (2003) J. Biol. Chem. 278: 9869 (e.g.5-methylpyrazole-3-carboxylic acid and acifran(4,5-dihydro-5-methyl-4-oxo-5-phenyl-2-furan carboxylic acidpyradine-3-acetic acid)), as well as 5-methyl nicotinic acid,nicotinuric acid, niceritrol, nicofuranose, acipimox(5-methylpyrazine-2-carboxylic acid 4-oxide), Niaspan® (niacinextended-release tablets; Kos) and those which can be easily identifiedby one skilled in the art which bind to and agonize the HM74A or HM74receptor (for example using the assays disclosed in Wise et al (2003) J.Biol. Chem 278:9869 (nicotine binding and [35S]-GTPyS binding assays),Soga et al (2003) Biochem. Biophys. Res. Comm. 303:364 (radiolabelbinding assay using the HM74 receptor which could be adapted to theHM74A receptor), Tunaru et al (2003) Nature Medicine 9:352 (calciummobilization assay using the HM74 receptor which could be adapted to theHM74A receptor) and U.S. Pat. No. 6,420,183 (FLIPR assays are describedgenerally in and may be adapted to the HM74A or HM74 receptor); reninangiotensin system inhibitors; bile acid reabsorption inhibitors (bileacid reuptake inhibitors), such as BARI 1453, SC435, PHA384640, S8921,AZD7706, and the like; PPARδ agonists (including partial agonists) suchas GW 501516, and GW 590735, and those disclosed in U.S. Pat. No.5,859,051 (acetophenols), WO03/024395, WO97/28149, WO01/79197,WO02/14291, WO02/46154, WO02/46176, WO02/076957, WO03/0 16291,WO03/033493, WO99/20275 (quinoline phenyl compounds), WO99/38845 (arylcompounds), WO00/63161 (1,4-disubstituted phenyl compounds), WO01/00579(aryl compounds), WO01/12612 & WO01/12187 (benzoic acid compounds), andWO97/31907 (substituted 4-hydroxy-phenylalconic acid compound); sterolbiosynthesis inhibitors such as DMP-565; triglyceride synthesisinhibitors; microsomal triglyceride transport (MTTP) inhibitors, such asinplitapide, LAB687, and CP346086, AEGR 733, implitapide and the like;HMG-CoA reductase gene expression inhibitors (e.g. compounds thatdecrease HMG-CoA reductase expression by affecting (e.g. blocking)transcription or translation of HMG-CoA reductase into protein orcompounds that maybe biotransformed into compounds that have theaforementioned attributes by one or more enzymes in the cholesterolbiosynthetic cascade or may lead to the accumulation of an isoprenemetabolite that has the aforementioned activities (such regulation isreadily determined by those skilled in the art according to standardassays (Methods of Enzymology, 110:9-19 1985))) such as those disclosedin U.S. Pat. No. 5,041,432 (certain 15-substituted lanosterolderivatives) and E. I. Mercer (1993) Prog. Lip. Res. 32:357 (oxygenatedsterols that suppress the biosynthesis of HMG-CoA reductase); squaleneepoxidase inhibitors such as NB-598((E)-N-ethyl-N-(6,6-dimethyl-2-hepten-4-y-nyl)-3-[(3,3′-bithiophen-5-yl)methoxy]benzene-methanaminehydrochloride); low density lipoprotein (LDL) receptor inducers such asHOE-402 (an imidazolidinyl-pyrimidine derivative that directlystimulates LDL receptor activity, see Huettinger et al (1993)Arterioscler. Thromb. 13:1005); platelet aggregation inhibitors; 5-LO orFLAP inhibitors; PPAR modulators (including compounds that may havemultiple functionality for activating various combinations of PPARα,PPARγ, and PPARδ) such as those disclosed in U.S. Pat. No. 6,008,237,U.S. Pat. No. 6,248,781, U.S. Pat. No. 6,166,049, WO00/12491,WO00/218355, WO00/23415, WO00/23416, WO00/23425, WO00/23442, WO00/23445,WO00/23451, WO00/236331, WO00/236332, WO00/238553, WO00/50392,WO00/53563, WO00/63153, WO00/63190, WO00/63196, WO00/63209, WO00/78312,WO00/78313, WO01/04351, WO01/14349, WO01/14350, WO01/16120, WO01/17994,WO01/21181, WO01/21578, WO01/25 181, WO01/25225, WO01/25226, WO01/40192,WO01/79150, WO02/081428, WO02/100403, WO02/102780, WO02/79162,WO03/016265, WO03/033453, WO03/042194, WO03/043997, WO03/066581,WO97/25042, WO99/07357, WO99/11255, WO99/12534, WO99/15520, WO99/46232,and WO98/05331 (including GW233 1 or (2-(4-[difluorophenyl]-1heptylureido)ethyl]phenoxy)-2-methylbutyric)); niacin-bound chromium, asdisclosed in WO03/039535; substituted acid derivatives disclosed inWO03/040114; apolipoprotein B inhibitors such as those disclosed inWO02/090347, WO02/28835, WO03/045921, WO03/047575; Factor Xa modulatorssuch as those disclosed in WO03/047517, WO03/047520, WO03/048081; ilealbile acid transport (“IBAT”) inhibitors (or apical sodium co-dependentbile acid transport (“ASBT”) inhibitors) such as benzothiepines(including 1,2-benzothiazepines; 1,4-benzodiazepines;1,5-benzothiazepines; 1,2, 5-benzothiadiazepines); PPARδ activators suchas disclosed in WO01/00603 (thiazole and oxazole derivates (e.g. C.A.S.Registry No. 317318-32-4), WO97/28149 (fluoro, chloro and thio phenoxyphenylacetic), U.S. Pat. No. 5,093,365 (non-1-oxidizable fatty acidanalogues), and WO99/04815. Tests showing the efficacy of the therapyand the rationale for the combination therapy with a dyslipidemic agentare presented in US2003 0069221 (where the dyslipidemic agents arecalled ‘cardiovascular agents’).

Dosage

Dosage levels of active ingredients in a pharmaceutical composition canalso be varied so as to achieve a transient or sustained concentrationof the compound in a subject, especially in and around the site ofinflammation or disease area, and to result in the desired response. Itis well within the skill of the art to start doses of the compound atlevels lower than required to achieve the desired effect and togradually increase the dosage until the desired effect is achieved. Itwill be understood that the specific dose level for any particularsubject will depend on a variety of factors, including body weight,general health, diet, natural history of disease, route and schedulingof administration, combination with one or more other drugs, andseverity of disease.

An effective dosage of the composition will typically be between about 1μg and about 10 mg per kilogram body weight, preferably between about 10μg to 5 mg of the compound per kilogram body weight. Adjustments indosage will be made using methods that are routine in the art and willbe based upon the particular composition being used and clinicalconsiderations.

The guanylate cyclase receptor agonists used in the methods describedabove may be administered orally, systemically or locally. Dosage formsinclude preparations for inhalation or injection, solutions,suspensions, emulsions, tablets, capsules, topical salves and lotions,transdermal compositions, other known peptide formulations and pegylatedpeptide analogs. Agonists may be administered as either the sole activeagent or in combination with other drugs, e.g., an inhibitor ofcGMP-dependent phosphodiesterase and anti-inflammatory agent. In allcases, additional drugs should be administered at a dosage that istherapeutically effective using the existing art as a guide. Drugs maybe administered in a single composition or sequentially.

Dosage levels of the Aad-GCRA peptides for use in methods of thisinvention typically are from about 0.001 mg to about 10,000 mg daily,preferably from about 0.005 mg to about 1,000 mg daily. For example, aneffective dosage of the Aad-GCRA peptides for use in methods of thisinvention is 0.1, 0.2. 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0,2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0,9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg per day oroptionally twice a day. Preferably the Aad-GCRA peptide is given after ameal (i.e, 30 minutes). In some embodiments a second agent describedabove is administered. In some aspects the second agent is administeredat less than the standard does for treating the particular disorderbecause the Aad-GCRA peptide acts synergistically with the second agent.For example, 2.5, 5. 7.5 or 10 mg of Liptor is given twice a day after ameal (i.e, 30 minutes). On the basis of mg/kg daily dose, either givenin single or divided doses, dosages typically range from about 0.001/75mg/kg to about 10,000/75 mg/kg, preferably from about 0.005/75 mg/kg toabout 1,000/75 mg/kg.

The total daily dose of each inhibitor can be administered to thepatient in a single dose, or in multiple subdoses. Typically, subdosescan be administered two to six times per day, preferably two to fourtimes per day, and even more preferably two to three times per day.Doses can be in immediate release form or sustained release formsufficiently effective to obtain the desired control over the medicalcondition.

The dosage regimen to prevent, treat, give relief from, or ameliorate amedical condition or disorder, or to otherwise protect against or treata medical condition with the combinations and compositions of thepresent invention is selected in accordance with a variety of factors.These factors include, but are not limited to, the type, age, weight,sex, diet, and medical condition of the subject, the severity of thedisease, the route of administration, pharmacological considerationssuch as the activity, efficacy, pharmacokinetics and toxicology profilesof the particular inhibitors employed, whether a drug delivery system isutilized, and whether the inhibitors are administered with other activeingredients. Thus, the dosage regimen actually employed may vary widelyand therefore deviate from the preferred dosage regimen set forth above.

I claim:
 1. A method for treating a condition that responds to enhancedcGMP levels in a patient comprising administering to said patient atherapeutically effective amount of a peptide consisting essentially ofthe sequence of any one of SEQ ID NOs: 1-38, 50-94, 119, and 120,wherein said peptide is administered in an amount sufficient to increasewater transport in the gastrointestinal tract and induce cGMP productionin a gastrointestinal epithelial cell.
 2. A method for preventing ortreating a condition selected from the group consisting of ulcerativecolitis, Crohn's disease, irritable bowel syndrome (IBS), non-ulcerdyspepsia, chronic intestinal pseudo-obstruction, functional dyspepsia,colonic pseudo-obstruction, duodenogastric reflux, constipation, chronicconstipation, constipation associated with use of opiate pain killers,post-surgical constipation, constipation associated with neuropathicdisorders, gastroesophageal reflux disease (GERD), Celiac disease,gastroparesis, heartburn, poor gastrointestinal motility, congestiveheart failure, hypertension, benign prostatic hyperplasia (BPH), coloncancer, lung cancer, bladder cancer, liver cancer, salivary gland canceror skin cancer, bronchitis, tissue inflammation, organ inflammation,respiratory inflammation, asthma, COPD, lipid metabolism disorders,biliary disorders, cardiovascular disease, obesity and an endocrinedisorder comprising administering to a patient in need thereof atherapeutically effective amount of a peptide consisting essentially ofthe sequence of any one of SEQ ID NOs: 1-38, 50-94, 119, and
 120. 3. Amethod of colonic cleansing, comprising administering to a subject inneed thereof an effective amount of a peptide consisting essentially ofthe sequence of any one of SEQ ID NOs: 1-38, 50-94, 119, and
 120. 4. Amethod of increasing cGMP production in a cell comprising contactingsaid cell with a peptide consisting essentially of the sequence of anyone of SEQ ID NOs: 1-38, 50-94, 119, and
 120. 5. The method of claim 1,wherein said peptide isAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶(SEQ ID NO: 1),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Ser¹⁶(SEQ ID NO: 32),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Thr¹⁶(SEQ ID NO: 119),Asn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Tyr¹⁶(SEQ ID NO: 120),dAsn¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-dLeu¹⁶(SEQ ID NO: 17), orpyGlu¹-Asp²-Aad³-Cys⁴-Glu⁵-Leu⁶-Cys⁷-Val⁸-Asn⁹-Val¹⁰-Ala¹¹-Cys¹²-Thr¹³-Gly¹⁴-Cys¹⁵-Leu¹⁶(SEQ ID NO: 56).
 6. The method of claim 1, further comprisingadministering a therapeutically effective amount of a cGMP-dependentphosphodiesterase inhibitor.
 7. The method of claim 6, wherein saidcGMP-dependent phosphodiesterase inhibitor is administered eitherconcurrently or sequentially with said guanylate cyclase receptoragonist.
 8. The method of claim 6, wherein said cGMP-dependentphosphodiesterase inhibitor is selected from the group consisting ofsulindac sulfone, zaprinast, motapizone, vardenafil, and sildenafil. 9.The method of claim 1, further comprising administering atherapeutically effective amount of at least one anti-inflammatoryagent.
 10. The method of 9, wherein said anti-inflammatory agent is asteroid or nonsteroid anti-inflammatory drug (NSAID).
 11. The method ofclaim 3, further comprising administering to said subject an effectiveamount of L-glucose, lubiprostone (Amitiza), prucalopride, an agent fortreating chronic constipation, or any combination thereof.